Quinazoline based egfr inhibitors containing a zinc binding moiety

ABSTRACT

The present invention relates to quinazoline containing zinc-binding moiety based derivatives that have enhanced and unexpected properties as inhibitors of epidermal growth factor receptor tyrosine kinase (EGFR-TK) and their use in the treatment of EGFR-TK related diseases and disorders such as cancer. The said derivatives may further act as HDAC inhibitors.

RELATED APPLICATIONS SECTION

This application is a continuation of U.S. application Ser. No.11/852,585, filed Sep. 10, 2007, which claims the benefit of U.S.Provisional Application No. 60/843,644, filed on Sep. 11, 2006 and U.S.Provisional Application No. 60/895,873, filed on Mar. 20, 2007, thecontents of which are hereby incorporated by reference. This applicationis also related to U.S. application Ser. No. 11/852,604, filed on Sep.10, 2007.

BACKGROUND OF THE INVENTION

The epidermal growth factor receptor (EGFR, Erb-B1) belongs to a familyof proteins, involved in the proliferation of normal and malignant cells(Artega, C. L., J. Clin Oncol 19, 2001, 32-40). Overexpression ofEpidermal Growth Factor Receptor (EGFR) is present in at least 70% ofhuman cancers (Seymour, L. K., Curr Drug Targets 2, 2001, 117-133) suchas, non-small cell lung carcinomas (NSCLC), breast cancers, gliomas,squamous cell carcinoma of the head and neck, and prostate cancer(Raymond et al., Drugs 60 Suppl 1, 2000, discussion 41-2; Salomon etal., Crit Rev Oncol Hematol 19, 1995, 183-232; Voldborg et al., AnnOncol 8, 1997, 1197-1206). The EGFR-TK is therefore widely recognized asan attractive target for the design and development of compounds thatcan specifically bind and inhibit the tyrosine kinase activity and itssignal transduction pathway in cancer cells, and thus can serve aseither diagnostic or therapeutic agents. For example, the EGFR tyrosinekinase (EGFR-TK) reversible inhibitor, TARCEVA®, was recently approvedby the FDA for treatment of NSCLC and advanced pancreatic cancer. Otheranti-EGFR targeted molecules have also been approved such as IRESSA®.

Despite the early success of Tarceva, it has become clear thatselectively targeting individual kinases can lead to the development ofdrug resistant tumors. Cells that have developed mutations within thedrug/kinase binding pocket display a growth advantage in the presence ofdrug eventually leading to disease progression. Current clinicalstrategies aimed at combining these molecularly targeted drugs withstandard chemotherapeutics, radiation, or other targeted agents willlead to novel strategies to improve overall response rate and increasethe number of complete remissions.

Furthermore, elucidation of the complex and multifactorial nature ofvarious diseases that involve multiple pathogenic pathways and numerousmolecular components suggests that multi-targeted therapies may beadvantageous over mono-therapies. Recent combination therapies with twoor more agents for many such diseases in the areas of oncology,infectious disease, cardiovascular disease and other complex pathologiesdemonstrate that this combinatorial approach may provide advantages withrespect to overcoming drug resistance, reduced toxicity and, in somecircumstances, a synergistic therapeutic effect compared to theindividual components.

Certain cancers have been effectively treated with such a combinatorialapproach; however, treatment regimes using a cocktail of cytotoxic drugsoften are limited by dose limiting toxicities and drug-druginteractions. More recent advances with molecularly targeted drugs haveprovided new approaches to combination treatment for cancer, allowingmultiple targeted agents to be used simultaneously, or combining thesenew therapies with standard chemotherapeutics or radiation to improveoutcome without reaching dose limiting toxicities. However, the abilityto use such combinations currently is limited to drugs that showcompatible pharmacologic and pharmacodynamic properties. In addition,the regulatory requirements to demonstrate safety and efficacy ofcombination therapies can be more costly and lengthy than correspondingsingle agent trials. Once approved, combination strategies may also beassociated with increased costs to patients, as well as decreasedpatient compliance owing to the more intricate dosing paradigmsrequired.

In the field of protein and polypeptide-based therapeutics it has becomecommonplace to prepare conjugates or fusion proteins that contain mostor all of the amino acid sequences of two differentproteins/polypeptides and that retain the individual binding activitiesof the separate proteins/polypeptides. This approach is made possible byindependent folding of the component protein domains and the large sizeof the conjugates that permits the components to bind their cellulartargets in an essentially independent manner. Such an approach is not,however, generally feasible in the case of small molecule therapeutics,where even minor structural modifications can lead to major changes intarget binding and/or the pharmacokinetic/pharmacodynamic properties ofthe resulting molecule.

The use of EGFR inhibitors in combination with histone deacetylases(HDAC) has been shown to produce synergistic effects. Histoneacetylation is a reversible modification, with deacetylation beingcatalyzed by a family of enzymes termed HDAC's. HDAC's are representedby X genes in humans and are divided into four distinct classes (J MolBiol, 2004, 338:1, 17-31). In mammalians class I HDAC's (HDAC1-3, andHDAC8) are related to yeast RPD3 HDAC, class 2 (HDAC4-7, HDAC9 andHDAC10) related to yeast HDA1, class 4 (HDAC11), and class 3 (a distinctclass encompassing the sirtuins which are related to yeast Sir2).

Csordas, Biochem. J., 1990, 286: 23-38 teaches that histones are subjectto post-translational acetylation of the, ε-amino groups of N-terminallysine residues, a reaction that is catalyzed by histone acetyltransferase (HAT1). Acetylation neutralizes the positive charge of thelysine side chain, and is thought to impact chromatin structure. Indeed,access of transcription factors to chromatin templates is enhanced byhistone hyperacetylation, and enrichment in underacetylated histone H4has been found in transcriptionally silent regions of the genome(Taunton et al., Science, 1996, 272:408-411). In the case of tumorsuppressor genes, transcriptional silencing due to histone modificationcan lead to oncogenic transformation and cancer.

Several classes of HDAC inhibitors currently are being evaluated byclinical investigators. The first FDA approved HDAC inhibitor isSuberoylanilide hydroxamic acid (SAHA, Zolinza®) for the treatment ofcutaneous T-cell lymphoma (CTCL). Other HDAC inhibitors includehydroxamic acid derivatives; PXD101 and LAQ824, are currently in theclinical development. In the benzamide class of HDAC inhibitors, MS-275,MGCD0103 and CI-994 have reached clinical trials. Mourne et al.(Abstract #4725, AACR 2005), demonstrate that thiophenyl modification ofbenzamides significantly enhance HDAC inhibitory activity against HDAC1.

Recent advances suggest that EGFR-TK inhibitors in combination with HDACinhibitors may provide advantageous results in the treatment of cancer.For example, co-treatment with SAHA significantly increased EGFR2antibody trastuzumab-induced apoptosis of BT-474 and SKBR-3 cells andinduced synergistic cytotoxic effects against the breast cancer cells(Bali, Clin. Cancer Res., 2005, 11, 3392). HDAC inhibitors, such asSAHA, have demonstrated synergistic antiproliferative and apoptoticeffects when used in combination with gefitinib in head and neck cancercell lines, including lines that are resistant to gefitinib monotherapy(Bruzzese et al., Proc. AACR, 2004). Pretreating gefitinib resistantcell lines with the HDAC inhibitor, MS-275, led to a growth-inhibitoryand apoptotic effect of gefitinib similar to that seen ingefitinib-sensitive NSCLC cell lines including those harboring EGFRmutations (Witta S. E., et al., Cancer Res 66:2, 2006, 944-50). The HDACinhibitor PXD101 has been shown to act synergistically to inhibitproliferation with the EGFR1 inhibitor Tarceva® (erlotinib)(WO2006082428A2).

Current therapeutic regimens of the types described above attempt toaddress the problem of drug resistance by the administration of multipleagents. However, the combined toxicity of multiple agents due tooff-target side effects as well as drug-drug interactions often limitsthe effectiveness of this approach. Moreover, it often is difficult tocombine compounds having differing pharmacokinetics into a single dosageform, and the consequent requirement of taking multiple medications atdifferent time intervals leads to problems with patient compliance thatcan undermine the efficacy of the drug combinations. In addition, thehealth care costs of combination therapies may be greater than the costof single molecule therapies. Furthermore, it may be more difficult toobtain regulatory approval of a combination therapy since the burden fordemonstrating activity/safety of a combination of two agents may begreater than for a single agent (Dancey J & Chen H, Nat. Rev. Drug Dis.,2006, 5:649). The development of novel agents that target multipletherapeutic targets selected not by virtue of cross reactivity, butthrough rational design will help improve patient outcome while avoidingthese limitations. Thus, enormous efforts are still directed to thedevelopment of selective anti-cancer drugs as well as to new and moreefficacious combinations of known anti-cancer drugs.

SUMMARY OF THE INVENTION

The present invention relates to quinazolines containing zinc-bindingmoiety based derivatives that have enhanced and unexpected properties asinhibitors of epidermal growth factor receptor tyrosine kinase(EGFR-TK). The compounds of the present invention also act as HDAC ormatrix metalloproteinase (MMP) inhibitors and HER2 inhibitors.Surprisingly, these compounds are active at multiple therapeutic targetsand are effective for treating diseases related to EGFR-TK activity,HDAC activity and/or HER2 activity, such as cancer and proliferativediseases. Moreover, it has even more surprisingly been found that thecompounds have enhanced activity when compared to the activities ofseparate molecules individually having the EGFR-TK and HDAC activitiesand combinations thereof. In other words, the combination ofpharmacophores into a single molecule may provide a synergistic effectas compared to the individual pharmacophores. More specifically, it hasbeen found that it is possible to prepare compounds that simultaneouslycontain a first portion of the molecule that binds zinc ions and thuspermits inhibition of HDAC and/or matrix metalloproteinase (MMP)activity and at least a second portion of the molecule that permitsbinding to a separate and distinct target that inhibits EGFR-TK and thusprovide therapeutic benefit. Preferably, the compounds of the presentinvention inhibit EGFR-TK, HER2 and HDAC activity.

Accordingly, the present invention provides a compound having thegeneral Formula I:

or its geometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof,wherein

-   -   X is O, S, CH₂, or —CONH—, preferably O;    -   B is a C₃ to C₉ alkylene, preferably a straight chain C₅ to C₇        alkylene, most preferably a straight chain C₆ alkylene;    -   R₁ is independently selected from hydrogen; C₁ to C₄ alkoxy,        preferably methoxy; or substituted C₁ to C₄ alkoxy, preferably        C₁ to C₄ alkoxy substituted C₁ to C₄ alkoxy, such as most        preferably methoxyethoxy; and    -   R₂ is each independently selected from halogen (preferably Br,        Cl and F), C₁ to C₄ alkyl, C₂ to C₄ alkenyl, and C₂ to C₄        alkynyl (preferably ethynyl);    -   n is 1, 2 or 3, preferably 1 or 2.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing and other objects, features and advantages of theinvention will be apparent from the following more particulardescription of preferred embodiments of the invention, as illustrated inthe accompanying drawings in which like reference characters refer tothe same parts throughout the different views. The drawings are notnecessarily to scale, emphasis instead being placed upon illustratingthe principles of the invention.

FIG. 1 (a) depicts a graph of EGFR enzyme assay results, (b) depicts agraph of HDAC enzyme assay results.

FIG. 2 illustrates inhibition of HDAC and EGFR in MDA-MB-468 breastcancer cell line: (a) Ac—H4 Accumulation, (b) Ac—H3 Accumulation, (c)EGFR inhibition.

FIG. 3 shows comparative data of anti-proliferative activity againstseveral different cancer cell lines: (a) pancreatic cancer (BxPC3), (b)NSCLC (H1703), (c) breast cancer (MDA-MB-468), (d) prostate cancer(PC3).

FIG. 4 illustrates the potency of compound 12 induction of apoptosis incancer cells: (a) HCT-116 (colon, 24 hours), (b) SKBr3 (breast, 24hours).

FIG. 5 shows the efficacy of compound 12 in A431 Epidermoid TumorXenograft Model (IP Dosing).

FIG. 6 shows the efficacy of compound 12 in H358 NSCLC Xenograft Model(2-Min IV infusion).

FIG. 7 shows the efficacy of compound 12 in H292 NSCLC Xenograft Model(2-Min IV infusion).

FIG. 8 shows the efficacy of compound 12 in BxPC3 Pancreatic CancerXenograft Model (2-Min IV infusion).

FIG. 9 shows the efficacy of compound 12 in PC3 Prostate CancerXenograft Model (2-Min IV infusion).

FIG. 10 shows the efficacy of compound 12 in HCT116 Colon CancerXenograft Model (2-Min IV infusion).

FIG. 11A shows the percent of change in tumor size in animals treatedwith compound 12 or vehicle in A549 NSCLC Xenograft model.

FIG. 11B shows the percent of change in tumor size in animals treatedwith Erlotinib and control in A549 NSCLC Xenograft model.

FIG. 12A shows the percent of change in tumor size in animals treatedwith compound 12, Erlotinib or vehicle in HPAC pancreatic cancer cells.

FIG. 12B shows the percent of change in body weight in animals treatedwith compound 12, Erlotinib or vehicle in HPAC pancreatic cancer cells.

FIG. 13 shows the concentration of compound 12 in plasma, lung and colonafter administration of hydrochloride, citrate, sodium and tartratesalts of compound 12.

FIG. 14 shows the concentration of compound 12 in the plasma of miceadministered compound 12 in 30% CAPTISOL.

FIG. 15 shows the percent change in mouse body weight afteradministration of an IV dose of compound 12 (25, 50, 100, 200 and 400mg/kg) in 30% CAPTISOL.

FIG. 16 shows the percent change in mouse body weight after 7 daysrepeat IP dosing of compound 12 (25, 50, 100, 200 and 400 mg/kg) in 30%CAPTISOL.

FIG. 17 shows the percent change in rat body weight after administrationof an IV dose of compound 12 (25, 50, 100 and 200 mg/kg) in 30%CAPTISOL.

DETAILED DESCRIPTION OF THE INVENTION

In a first embodiment of the compounds of the present invention arecompounds represented by formula (I) as illustrated above, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof.

In a second embodiment of the compounds of the present invention arecompounds represented by formula (II) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein B, R₁, R₂, and n are as previously defined.

In a third embodiment of the compounds of the present invention arecompounds represented by formula (ITT) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein B and R₁ are as previously defined.

In a fourth embodiment of the compounds of the present invention arecompounds represented by formula (IV) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein B and R₁ are as previously defined.

In each of the above embodiments, R₁ is preferably hydrogen or methoxyand, independently or collectively, B is preferably a straight chain C₅to C₇ alkylene, most preferably a straight chain C₆ alkylene.

Representative compounds according to the invention claimed herein, inthe priority applications of this application and in related applicationU.S. Ser. No. 11/852,604, corresponding to Attorney Docket No. 4045.3001US2, filed on even date herewith are those selected from the Table Abelow or its geometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

TABLE A Compound # Structure 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

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37

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41

42

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50

51

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62

63

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67

68

69

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71

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77

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79

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81

82

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86

87

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89

90

91

92

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94

95

96

97

98

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100

101

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

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138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

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158

159

160

161

162

163

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165

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167

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182

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184

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209

210

211

212

In a particularly preferred embodiment, the invention relates togeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates of compounds 12and 18.

The invention further provides methods for the prevention or treatmentof diseases or conditions involving aberrant proliferation,differentiation or survival of cells. In one embodiment, the inventionfurther provides for the use of one or more compounds of the inventionin the manufacture of a medicament for halting or decreasing diseasesinvolving aberrant proliferation, differentiation, or survival of cells.In preferred embodiments, the disease is cancer. In one embodiment, theinvention relates to a method of treating cancer in a subject in need oftreatment comprising administering to said subject a therapeuticallyeffective amount of a compound of the invention.

The term “cancer” refers to any cancer caused by the proliferation ofmalignant neoplastic cells, such as tumors, neoplasms, carcinomas,sarcomas, leukemias, lymphomas and the like. For example, cancersinclude, but are not limited to, mesothelioma, leukemias and lymphomassuch as cutaneous T-cell lymphomas (CTCL), noncutaneous peripheralT-cell lymphomas, lymphomas associated with human T-cell lymphotrophicvirus (HTLV) such as adult T-cell leukemia/lymphoma (ATLL), B-celllymphoma, acute nonlymphocytic leukemias, chronic lymphocytic leukemia,chronic myelogenous leukemia, acute myelogenous leukemia, lymphomas, andmultiple myeloma, non-Hodgkin lymphoma, acute lymphatic leukemia (ALL),chronic lymphatic leukemia (CLL), Hodgkin's lymphoma, Burkitt lymphoma,adult T-cell leukemia lymphoma, acute-myeloid leukemia (AML), chronicmyeloid leukemia (CML), or hepatocellular carcinoma. Further examplesinclude myelodisplastic syndrome, childhood solid tumors such as braintumors, neuroblastoma, retinoblastoma, Wilms' tumor, bone tumors, andsoft-tissue sarcomas, common solid tumors of adults such as head andneck cancers (e.g., oral, laryngeal, nasopharyngeal and esophageal),genitourinary cancers (e.g., prostate, bladder, renal, uterine, ovarian,testicular), lung cancer (e.g., small-cell and non small cell), breastcancer, pancreatic cancer, melanoma and other skin cancers, stomachcancer, brain tumors, tumors related to Gorlin's syndrome (e.g.,medulloblastoma, meningioma, etc.), and liver cancer. Additionalexemplary forms of cancer which may be treated by the subject compoundsinclude, but are not limited to, cancer of skeletal or smooth muscle,stomach cancer, cancer of the small intestine, rectum carcinoma, cancerof the salivary gland, endometrial cancer, adrenal cancer, anal cancer,rectal cancer, parathyroid cancer, and pituitary cancer.

Additional cancers that the compounds described herein may be useful inpreventing, treating and studying are, for example, colon carcinoma,familiary adenomatous polyposis carcinoma and hereditary non-polyposiscolorectal cancer, or melanoma. Further, cancers include, but are notlimited to, labial carcinoma, larynx carcinoma, hypopharynx carcinoma,tongue carcinoma, salivary gland carcinoma, gastric carcinoma,adenocarcinoma, thyroid cancer (medullary and papillary thyroidcarcinoma), renal carcinoma, kidney parenchyma carcinoma, cervixcarcinoma, uterine corpus carcinoma, endometrium carcinoma, chorioncarcinoma, testis carcinoma, urinary carcinoma, melanoma, brain tumorssuch as glioblastoma, astrocytoma, meningioma, medulloblastoma andperipheral neuroectodermal tumors, gall bladder carcinoma, bronchialcarcinoma, multiple myeloma, basalioma, teratoma, retinoblastoma,choroidea melanoma, seminoma, rhabdomyosarcoma, craniopharyngeoma,osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma,Ewing sarcoma, and plasmocytoma. In one aspect of the invention, thepresent invention provides for the use of one or more compounds of theinvention in the manufacture of a medicament for the treatment ofcancer.

In one embodiment, the present invention includes the use of one or morecompounds of the invention in the manufacture of a medicament thatprevents further aberrant proliferation, differentiation, or survival ofcells. For example, compounds of the invention may be useful inpreventing tumors from increasing in size or from reaching a metastaticstate. The subject compounds may be administered to halt the progressionor advancement of cancer or to induce tumor apoptosis or to inhibittumor angiogenesis. In addition, the instant invention includes use ofthe subject compounds to prevent a recurrence of cancer.

This invention further embraces the treatment or prevention of cellproliferative disorders such as hyperplasias, dysplasias andpre-cancerous lesions. Dysplasia is the earliest form of pre-cancerouslesion recognizable in a biopsy by a pathologist. The subject compoundsmay be administered for the purpose of preventing said hyperplasias,dysplasias or pre-cancerous lesions from continuing to expand or frombecoming cancerous. Examples of pre-cancerous lesions may occur in skin,esophageal tissue, breast and cervical intra-epithelial tissue.

“Combination therapy” includes the administration of the subjectcompounds in further combination with other biologically activeingredients (such as, but not limited to, a second and differentantineoplastic agent) and non-drug therapies (such as, but not limitedto, surgery or radiation treatment). For instance, the compounds of theinvention can be used in combination with other pharmaceutically activecompounds, preferably compounds that are able to enhance the effect ofthe compounds of the invention. The compounds of the invention can beadministered simultaneously (as a single preparation or separatepreparation) or sequentially to the other drug therapy. In general, acombination therapy envisions administration of two or more drugs duringa single cycle or course of therapy.

In one aspect of the invention, the subject compounds may beadministered in combination with one or more separate agents thatmodulate protein kinases involved in various disease states. Examples ofsuch kinases may include, but are not limited to: serine/threoninespecific kinases, receptor tyrosine specific kinases and non-receptortyrosine specific kinases. Serine/threonine kinases include mitogenactivated protein kinases (MAPK), meiosis specific kinase (MEK), RAF andaurora kinase. Examples of receptor kinase families include epidermalgrowth factor receptor (EGFR) (e.g. HER2/neu, HER3, HER4, ErbB, ErbB2,ErbB3, ErbB4, Xmrk, DER, Let23); fibroblast growth factor (FGF) receptor(e.g. FGF-R1, GFF-R2/BEK/CEK3, FGF-R3/CEK2, FGF-R4/TKF, KGF-R);hepatocyte growth/scatter factor receptor (HGFR) (e.g, MET, RON, SEA,SEX); insulin receptor (e.g. IGFI-R); Eph (e.g. CEK5, CEK8, EBK, ECK,EEK, EHK-1, EHK-2, ELK, EPH, ERK, HEK, MDK2, MDK5, SEK); Axl (e.g.Mer/Nyk, Rse); RET; and platelet-derived growth factor receptor (PDGFR)(e.g. PDGFα-R, PDGβ-R, CSF1-R/FMS, SCF-R/C-KIT, VEGF-R/FLT, NEK/FLK1,FLT3/FLK2/STK-1). Non-receptor tyrosine kinase families include, but arenot limited to, BCR-ABL (e.g. p43^(abl), ARG); BTK (e.g. ITK/EMT, TEC);CSK, FAK, FPS, JAK, SRC, BMX, FER, CDK and SYK.

In another aspect of the invention, the subject compounds may beadministered in combination with one or more separate agents thatmodulate non-kinase biological targets or processes. Such targetsinclude histone deacetylases (HDAC), DNA methyltransferase (DNMT), heatshock proteins (e.g. HSP90), and proteosomes.

In a preferred embodiment, subject compounds may be combined withantineoplastic agents (e.g. small molecules, monoclonal antibodies,antisense RNA, and fusion proteins) that inhibit one or more biologicaltargets such as Zolinza, Tarceva, Iressa, Tykerb, Gleevec, Sutent,Sprycel, Nexavar, Sorafinib, CNF2024, RG108, BMS387032, Affinitak,Avastin, Herceptin, Erbitux, AG24322, PD325901, ZD6474, PD184322,Obatodax, ABT737 and AEE788. Such combinations may enhance therapeuticefficacy over efficacy achieved by any of the agents alone and mayprevent or delay the appearance of resistant mutational variants.

In certain preferred embodiments, the compounds of the invention areadministered in combination with a chemotherapeutic agent.Chemotherapeutic agents encompass a wide range of therapeutic treatmentsin the field of oncology. These agents are administered at variousstages of the disease for the purposes of shrinking tumors, destroyingremaining cancer cells left over after surgery, inducing remission,maintaining remission and/or alleviating symptoms relating to the canceror its treatment. Examples of such agents include, but are not limitedto, alkylating agents such as mustard gas derivatives (Mechlorethamine,cylophosphamide, chlorambucil, melphalan, ifosfamide), ethylenimines(thiotepa, hexamethylmelanine), Alkylsulfonates (Busulfan), Hydrazinesand Triazines (Altretamine, Procarbazine, Dacarbazine and Temozolomide),Nitrosoureas (Carmustine, Lomustine and Streptozocin), Ifosfamide andmetal salts (Carboplatin, Cisplatin, and Oxaliplatin); plant alkaloidssuch as Podophyllotoxins (Etoposide and Tenisopide), Taxanes (Paclitaxeland Docetaxel), Vinca alkaloids (Vincristine, Vinblastine, Vindesine andVinorelbine), and Camptothecan analogs (Irinotecan and Topotecan);anti-tumor antibiotics such as Chromomycins (Dactinomycin andPlicamycin), Anthracyclines (Doxorubicin, Daunorubicin, Epirubicin,Mitoxantrone, Valrubicin and Idarubicin), and miscellaneous antibioticssuch as Mitomycin, Actinomycin and Bleomycin; anti-metabolites such asfolic acid antagonists (Methotrexate, Pemetrexed, Raltitrexed,Aminopterin), pyrimidine antagonists (5-Fluorouracil, Floxuridine,Cytarabine, Capecitabine, and Gemcitabine), purine antagonists(6-Mercaptopurine and 6-Thioguanine) and adenosine deaminase inhibitors(Cladribine, Fludarabine, Mercaptopurine, Clofarabine, Thioguanine,Nelarabine and Pentostatin); topoisomerase inhibitors such astopoisomerase I inhibitors (Ironotecan, topotecan) and topoisomerase IIinhibitors (Amsacrine, etoposide, etoposide phosphate, teniposide);monoclonal antibodies (Alemtuzumab, Gemtuzumab ozogamicin, Rituximab,Trastuzumab, Ibritumomab Tioxetan, Cetuximab, Panitumumab, Tositumomab,Bevacizumab); and miscellaneous anti-neoplastics such as ribonucleotidereductase inhibitors (Hydroxyurea); adrenocortical steroid inhibitor(Mitotane); enzymes (Asparaginase and Pegaspargase); anti-microtubuleagents (Estramustine); and retinoids (Bexarotene, Isotretinoin,Tretinoin (ATRA).

In certain preferred embodiments, the compounds of the invention areadministered in combination with a chemoprotective agent.Chemoprotective agents act to protect the body or minimize the sideeffects of chemotherapy. Examples of such agents include, but are notlimited to, amfostine, mesna, and dexrazoxane.

In one aspect of the invention, the subject compounds are administeredin combination with radiation therapy. Radiation is commonly deliveredinternally (implantation of radioactive material near cancer site) orexternally from a machine that employs photon (x-ray or gamma-ray) orparticle radiation. Where the combination therapy further comprisesradiation treatment, the radiation treatment may be conducted at anysuitable time so long as a beneficial effect from the co-action of thecombination of the therapeutic agents and radiation treatment isachieved. For example, in appropriate cases, the beneficial effect isstill achieved when the radiation treatment is temporally removed fromthe administration of the therapeutic agents, perhaps by days or evenweeks.

It will be appreciated that compounds of the invention can be used incombination with an immunotherapeutic agent. One form of immunotherapyis the generation of an active systemic tumor-specific immune responseof host origin by administering a vaccine composition at a site distantfrom the tumor. Various types of vaccines have been proposed, includingisolated tumor-antigen vaccines and anti-idiotype vaccines. Anotherapproach is to use tumor cells from the subject to be treated, or aderivative of such cells (reviewed by Schirrmacher et al. (1995) J.Cancer Res. Clin. Oncol. 121:487). In U.S. Pat. No. 5,484,596, Hanna Jr.et al. claim a method for treating a resectable carcinoma to preventrecurrence or metastases, comprising surgically removing the tumor,dispersing the cells with collagenase, irradiating the cells, andvaccinating the patient with at least three consecutive doses of about10⁷ cells.

It will be appreciated that the compounds of the invention mayadvantageously be used in conjunction with one or more adjunctivetherapeutic agents. Examples of suitable agents for adjunctive therapyinclude a 5HT₁ agonist, such as a triptan (e.g. sumatriptan ornaratriptan); an adenosine A1 agonist; an EP ligand; an NMDA modulator,such as a glycine antagonist; a sodium channel blocker (e.g.lamotrigine); a substance P antagonist (e.g. an NK₁ antagonist); acannabinoid; acetaminophen or phenacetin; a 5-lipoxygenase inhibitor; aleukotriene receptor antagonist; a DMARD (e.g. methotrexate); gabapentinand related compounds; a tricyclic antidepressant (e.g. amitryptilline);a neurone stabilising antiepileptic drug; a mono-aminergic uptakeinhibitor (e.g. venlafaxine); a matrix metalloproteinase inhibitor; anitric oxide synthase (NOS) inhibitor, such as an iNOS or an nNOSinhibitor; an inhibitor of the release, or action, of tumour necrosisfactor .alpha.; an antibody therapy, such as a monoclonal antibodytherapy; an antiviral agent, such as a nucleoside inhibitor (e.g.lamivudine) or an immune system modulator (e.g. interferon); an opioidanalgesic; a local anaesthetic; a stimulant, including caffeine; anH₂-antagonist (e.g. ranitidine); a proton pump inhibitor (e.g.omeprazole); an antacid (e.g. aluminium or magnesium hydroxide; anantiflatulent (e.g. simethicone); a decongestant (e.g. phenylephrine,phenylpropanolamine, pseudoephedrine, oxymetazoline, epinephrine,naphazoline, xylometazoline, propylhexedrine, or levo-desoxyephedrine);an antitussive (e.g. codeine, hydrocodone, carmiphen, carbetapentane, ordextramethorphan); a diuretic; or a sedating or non-sedatingantihistamine.

Matrix metalloproteinases (MMPs) are a family of zinc-dependent neutralendopeptidases collectively capable of degrading essentially all matrixcomponents. Over 20 MMP modulating agents are in pharmaceutical develop,almost half of which are indicated for cancer. The University of Torontoresearchers have reported that HDACs regulate MMP expression andactivity in 3T3 cells. In particular, inhibition of HDAC by trichostatinA (TSA), which has been shown to prevent tumorigenesis and metastasis,decreases mRNA as well as zymographic activity of gelatinase A (MMP2;Type IV collagenase), a matrix metalloproteinase, which is itself,implicated in tumorigenesis and metastasis (Ailenberg M., Silverman M.,Biochem Biophys Res Commun. 2002, 298:110-115). Another recent articlethat discusses the relationship of HDAC and MMPs can be found in YoungD. A., et al., Arthritis Research & Therapy, 2005, 7: 503. Furthermore,the commonality between HDAC and MMPs inhibitors is their zinc-bindingfunctionality. Therefore, in one aspect of the invention, compounds ofthe invention can be used as MMP inhibitors and may be of use in thetreatment of disorders relating to or associated with dysregulation ofMMP. The overexpression and activation of MMPs are known to inducetissue destruction and are also associated with a number of specificdiseases including rheumatoid arthritis, periodontal disease, cancer andatherosclerosis.

The compounds may also be used in the treatment of a disorder involving,relating to or, associated with dysregulation of histone deacetylase(HDAC). There are a number of disorders that have been implicated by orknown to be mediated at least in part by HDAC activity, where HDACactivity is known to play a role in triggering disease onset, or whosesymptoms are known or have been shown to be alleviated by HDACinhibitors. Disorders of this type that would be expected to be amenableto treatment with the compounds of the invention include the followingbut not limited to: Anti-proliferative disorders (e.g. cancers);Neurodegenerative diseases including Huntington's Disease, Polyglutaminedisease, Parkinson's Disease, Alzheimer's Disease, Seizures,Striatonigral degeneration, Progressive supranuclear palsy, Torsiondystonia, Spasmodic torticollis and dyskinesis, Familial tremor, Gillesde la Tourette syndrome, Diffuse Lewy body disease, Progressivesupranuclear palsy, Pick's disease, intracerebral hemorrhage, Primarylateral sclerosis, Spinal muscular atrophy, Amyotrophic lateralsclerosis, Hypertrophic interstitial polyneuropathy, Retinitispigmentosa, Hereditary optic atrophy, Hereditary spastic paraplegia,Progressive ataxia and Shy-Drager syndrome; Metabolic diseases includingType 2 diabetes; Degenerative Diseases of the Eye including Glaucoma,Age-related macular degeneration, Rubeotic glaucoma; Inflammatorydiseases and/or Immune system disorders including Rheumatoid Arthritis(RA), Osteoarthritis, Juvenile chronic arthritis, Graft versus Hostdisease, Psoriasis, Asthma, Spondyloarthropathy, Crohn's Disease,inflammatory bowel disease Colitis Ulcerosa, Alcoholic hepatitis,Diabetes, Sjoegrens's syndrome, Multiple Sclerosis, Ankylosingspondylitis, Membranous glomerulopathy, Discogenic pain, Systemic LupusErythematosus; Disease involving angiogenesis including cancer,psoriasis, rheumatoid arthritis; Psychological disorders includingbipolar disease, schizophrenia, mania, depression and dementia;Cardiovascular Diseases including heart failure, restenosis andarteriosclerosis; Fibrotic diseases including liver fibrosis, cysticfibrosis and angiofibroma; Infectious diseases including Fungalinfections, such as Candida Albicans, Bacterial infections, Viralinfections, such as Herpes Simplex, Protozoal infections, such asMalaria, Leishmania infection, Trypanosoma brucei infection,Toxoplasmosis and coccidlosis and Haematopoietic disorders includingthalassemia, anemia and sickle cell anemia.

In one embodiment, compounds of the invention can be used to induce orinhibit apoptosis, a physiological cell death process critical fornormal development and homeostasis. Alterations of apoptotic pathwayscontribute to the pathogenesis of a variety of human diseases. Compoundsof the invention, as modulators of apoptosis, will be useful in thetreatment of a variety of human diseases with abberations in apoptosisincluding cancer (particularly, but not limited to, follicularlymphomas, carcinomas with p53 mutations, hormone dependent tumors ofthe breast, prostate and ovary, and precancerous lesions such asfamilial adenomatous polyposis), viral infections (including, but notlimited to, herpesvirus, poxvirus, Epstein-Barr virus, Sindbis virus andadenovirus), autoimmune diseases (including, but not limited to,systemic lupus, erythematosus, immune mediated glomerulonephritis,rheumatoid arthritis, psoriasis, inflammatory bowel diseases, andautoimmune diabetes mellitus), neurodegenerative disorders (including,but not limited to, Alzheimer's disease, AIDS-related dementia,Parkinson's disease, amyotrophic lateral sclerosis, retinitispigmentosa, spinal muscular atrophy and cerebellar degeneration), AIDS,myelodysplastic syndromes, aplastic anemia, ischemic injury associatedmyocardial infarctions, stroke and reperfusion injury, arrhythmia,atherosclerosis, toxin-induced or alcohol induced liver diseases,hematological diseases (including, but not limited to, chronic anemiaand aplastic anemia), degenerative diseases of the musculoskeletalsystem (including, but not limited to, osteoporosis and arthritis),aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple sclerosis,kidney diseases, and cancer pain.

In one aspect, the invention provides the use of compounds of theinvention for the treatment and/or prevention of immune response orimmune-mediated responses and diseases, such as the prevention ortreatment of rejection following transplantation of synthetic or organicgrafting materials, cells, organs or tissue to replace all or part ofthe function of tissues, such as heart, kidney, liver, bone marrow,skin, cornea, vessels, lung, pancreas, intestine, limb, muscle, nervetissue, duodenum, small-bowel, pancreatic-islet-cell, includingxeno-transplants, etc.; to treat or prevent graft-versus-host disease,autoimmune diseases, such as rheumatoid arthritis, systemic lupuserythematosus, thyroiditis, Hashimoto's thyroiditis, multiple sclerosis,myasthenia gravis, type I diabetes uveitis, juvenile-onset orrecent-onset diabetes mellitus, uveitis, Graves disease, psoriasis,atopic dermatitis, Crohn's disease, ulcerative colitis, vasculitis,auto-antibody mediated diseases, aplastic anemia, Evan's syndrome,autoimmune hemolytic anemia, and the like; and further to treatinfectious diseases causing aberrant immune response and/or activation,such as traumatic or pathogen induced immune disregulation, includingfor example, that which are caused by hepatitis B and C infections, HIV,staphylococcus aureus infection, viral encephalitis, sepsis, parasiticdiseases wherein damage is induced by an inflammatory response (e.g.,leprosy); and to prevent or treat circulatory diseases, such asarteriosclerosis, atherosclerosis, vasculitis, polyarteritis nodosa andmyocarditis. In addition, the present invention may be used toprevent/suppress an immune response associated with a gene therapytreatment, such as the introduction of foreign genes into autologouscells and expression of the encoded product. Thus in one embodiment, theinvention relates to a method of treating an immune response disease ordisorder or an immune-mediated response or disorder in a subject in needof treatment comprising administering to said subject a therapeuticallyeffective amount of a compound of the invention.

In one aspect, the invention provides the use of compounds of theinvention in the treatment of a variety of neurodegenerative diseases, anon-exhaustive list of which includes: I. Disorders characterized byprogressive dementia in the absence of other prominent neurologic signs,such as Alzheimer's disease; Senile dementia of the Alzheimer type; andPick's disease (lobar atrophy); II. Syndromes combining progressivedementia with other prominent neurologic abnormalities such as A)syndromes appearing mainly in adults (e.g., Huntington's disease,Multiple system atrophy combining dementia with ataxia and/ormanifestations of Parkinson's disease, Progressive supranuclear palsy(Steel-Richardson-Olszewski), diffuse Lewy body disease, andcorticodentatonigral degeneration); and B) syndromes appearing mainly inchildren or young adults (e.g., Hallervorden-Spatz disease andprogressive familial myoclonic epilepsy); III. Syndromes of graduallydeveloping abnormalities of posture and movement such as paralysisagitans (Parkinson's disease), striatonigral degeneration, progressivesupranuclear palsy, torsion dystonia (torsion spasm; dystonia musculorumdeformans), spasmodic torticollis and other dyskinesis, familial tremor,and Gilles de la Tourette syndrome; IV. Syndromes of progressive ataxiasuch as cerebellar degenerations (e.g., cerebellar cortical degenerationand olivopontocerebellar atrophy (OPCA)); and spinocerebellardegeneration (Friedreich's atazia and related disorders); V. Syndrome ofcentral autonomic nervous system failure (Shy-Drager syndrome); VI.Syndromes of muscular weakness and wasting without sensory changes(motorneuron disease such as amyotrophic lateral sclerosis, spinalmuscular atrophy (e.g., infantile spinal muscular atrophy(Werdnig-Hoffman), juvenile spinal muscular atrophy(Wohlfart-Kugelberg-Welander) and other forms of familial spinalmuscular atrophy), primary lateral sclerosis, and hereditary spasticparaplegia; VII. Syndromes combining muscular weakness and wasting withsensory changes (progressive neural muscular atrophy; chronic familialpolyneuropathies) such as peroneal muscular atrophy(Charcot-Marie-Tooth), hypertrophic interstitial polyneuropathy(Dejerine-Sottas), and miscellaneous forms of chronic progressiveneuropathy; VIII Syndromes of progressive visual loss such as pigmentarydegeneration of the retina (retinitis pigmentosa), and hereditary opticatrophy (Leber's disease). Furthermore, compounds of the invention canbe implicated in chromatin remodeling.

The invention encompasses pharmaceutical compositions comprisingpharmaceutically acceptable salts of or complexes with the compounds ofthe invention as described above. Examples of suitable salts orcomplexes include but are not limited to the sodium, hydrochloride,citrate or tartrate salt, preferably the tartrate salt. The inventionalso encompasses pharmaceutical compositions comprising solvates orhydrates of the compounds of the invention. The term “hydrate” includesbut is not limited to hemihydrate, monohydrate, dihydrate, trihydrateand the like. In yet another embodiment, the invention relates to atartrate salt of or complex with compound 12. In an additionalembodiment, the invention is an L-tartrate salt of or complex withcompound 12.

The tartrate salts or complexes of the invention include the L-tartrate(L-(R,R)-(+)-tartrate), D-tartrate (D-((S,S)-(−)-tartrate), D,L-tartrateand meso-tartrate (2R,3S-tartrate) salts. In another embodiment, thetartrate salt is an anhydrous salt. In a further embodiment, thetartrate salt is a hydrate salt. The term “hydrate” includes but is notlimited to hemihydrate, monohydrate, dihydrate, trihydrate and the like.

In a further embodiment, the invention pertains to an L-tartrate salt ofor complex with a compound represented by Formulae I, II, III or IV. Inyet another embodiment, the invention is an L-tartrate salt of orcomplex with a compound selected from the compounds 1-197 shown in TableA above. In an additional embodiment, the invention is an L-tartratesalt of or complex with a compound selected from the group consisting ofcompound 12 and compound 18. In a further embodiment, the invention isan L-tartrate salt of or complex with compound 12.

In an additional embodiment, the invention pertains to a D-tartrate saltof or complex with a compound represented by Formulae I, II, III or IV.In yet another embodiment, the invention is a D-tartrate salt of orcomplex with a compound selected from the compounds 1-197 shown in TableA above. In an additional embodiment, the invention is a D-tartrate saltof or complex with a compound selected from the group consisting ofcompound 12 and compound 18. In a further embodiment, the invention is aD-tartrate salt of or complex with compound 12.

In a further embodiment, the invention pertains to an D,L-tartrate saltof or complex with a compound represented by Formulae I, II, III or IV.In yet another embodiment, the invention is an D,L-tartrate salt of orcomplex with a compound selected from the compounds 1-197 shown in TableA above. In an additional embodiment, the invention is an D,L-tartratesalt of or complex with a compound selected from the group consisting ofcompound 12 and compound 18. In a further embodiment, the invention isan D,L-tartrate salt of or complex with compound 12.

In yet another embodiment, the invention is a meso-tartrate salt of orcomplex with a compound represented by Formulae I, II, III or IV. In yetanother embodiment, the invention is a meso-tartrate salt of or complexwith a compound selected from the compounds 1-197 shown in Table Aabove. In an additional embodiment, the invention is a meso-tartratesalt of or complex with a compound selected from the group consisting ofcompound 12 and compound 18. In a further embodiment, the invention is ameso-tartrate salt of or complex with compound 12.

In one embodiment, the invention pertains to a pharmaceuticalcomposition comprising a tartrate salt of or complex with the inventionat a therapeutically effective amount and a pharmaceutically acceptablecarrier. In another embodiment, the pharmaceutical composition is aliquid formulation. In yet another embodiment, the pharmaceuticalcomposition is an aqueous formulation. In yet another embodiment, theliquid formulation is a parenteral formulation. In yet anotherembodiment, the formulation is an intravenous formulation. The inventionfurther encompasses pharmaceutical compositions comprising any solid orliquid physical form of the compound of the invention. The particles maybe micronized, or may be agglomerated, particulate granules, powders,oils, oily suspensions or any other form of solid or liquid physicalform.

The compounds of the invention, and derivatives, fragments, analogs,homologs, pharmaceutically acceptable salts or hydrate thereof can beincorporated into pharmaceutical compositions suitable foradministration, together with a pharmaceutically acceptable carrier orexcipient. Such compositions typically comprise a therapeuticallyeffective amount of any of the compounds above, and a pharmaceuticallyacceptable carrier. Preferably, the effective amount when treatingcancer is an amount effective to selectively induce terminaldifferentiation of suitable neoplastic cells and less than an amountwhich causes toxicity in a patient.

Compounds of the invention may be administered by any suitable means,including, without limitation, parenteral, intravenous, intramuscular,subcutaneous, implantation, oral, sublingual, buccal, nasal, pulmonary,transdermal, topical, vaginal, rectal, and transmucosal administrationsor the like. Topical administration can also involve the use oftransdermal administration such as transdermal patches or iontophoresisdevices. Pharmaceutical preparations include a solid, semisolid orliquid preparation (tablet, pellet, troche, capsule, suppository, cream,ointment, aerosol, powder, liquid, emulsion, suspension, syrup,injection etc.) containing a compound of the invention as an activeingredient, which is suitable for selected mode of administration. Inone embodiment, the pharmaceutical compositions are administered orally,and are thus formulated in a form suitable for oral administration,i.e., as a solid or a liquid preparation. Suitable solid oralformulations include tablets, capsules, pills, granules, pellets,sachets and effervescent, powders, and the like. Suitable liquid oralformulations include solutions, suspensions, dispersions, emulsions,oils and the like. In one embodiment of the present invention, thecomposition is formulated in a capsule. In accordance with thisembodiment, the compositions of the present invention comprise inaddition to the active compound and the inert carrier or diluent, a hardgelatin capsule.

Any inert excipient that is commonly used as a carrier or diluent may beused in the formulations of the present invention, such as for example,a gum, a starch, a sugar, a cellulosic material, an acrylate, ormixtures thereof. A preferred diluent is microcrystalline cellulose. Thecompositions may further comprise a disintegrating agent (e.g.,croscarmellose sodium) and a lubricant (e.g., magnesium stearate), andmay additionally comprise one or more additives selected from a binder,a buffer, a protease inhibitor, a surfactant, a solubilizing agent, aplasticizer, an emulsifier, a stabilizing agent, a viscosity increasingagent, a sweetener, a film forming agent, or any combination thereof.Furthermore, the compositions of the present invention may be in theform of controlled release or immediate release formulations.

For liquid formulations, pharmaceutically acceptable carriers may beaqueous or non-aqueous solutions, suspensions, emulsions or oils.Examples of non-aqueous solvents are propylene glycol, polyethyleneglycol, and injectable organic esters such as ethyl oleate. Aqueouscarriers include water, alcoholic/aqueous solutions, emulsions orsuspensions, including saline and buffered media. Examples of oils arethose of petroleum, animal, vegetable, or synthetic origin, for example,peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, andfish-liver oil. Solutions or suspensions can also include the followingcomponents: a sterile diluent such as water for injection, salinesolution, fixed oils, polyethylene glycols, glycerine, propylene glycolor other synthetic solvents; antibacterial agents such as benzyl alcoholor methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid(EDTA); buffers such as acetates, citrates or phosphates, and agents forthe adjustment of tonicity such as sodium chloride or dextrose. The pHcan be adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide.

In addition, the compositions may further comprise binders (e.g.,acacia, cornstarch, gelatin, carbomer, ethyl cellulose, guar gum,hydroxypropyl cellulose, hydroxypropyl methyl cellulose, povidone),disintegrating agents (e.g., cornstarch, potato starch, alginic acid,silicon dioxide, croscarmellose sodium, crospovidone, guar gum, sodiumstarch glycolate, Primogel), buffers (e.g., tris-HCI., acetate,phosphate) of various pH and ionic strength, additives such as albuminor gelatin to prevent absorption to surfaces, detergents (e.g., Tween20, Tween 80, Pluronic F68, bile acid salts), protease inhibitors,surfactants (e.g., sodium lauryl sulfate), permeation enhancers,solubilizing agents (e.g., glycerol, polyethylene glycerol), a glidant(e.g., colloidal silicon dioxide), anti-oxidants (e.g., ascorbic acid,sodium metabisulfite, butylated hydroxyanisole), stabilizers (e.g.,hydroxypropyl cellulose, hydroxypropylmethyl cellulose), viscosityincreasing agents (e.g., carbomer, colloidal silicon dioxide, ethylcellulose, guar gum), sweeteners (e.g., sucrose, aspartame, citricacid), flavoring agents (e.g., peppermint, methyl salicylate, or orangeflavoring), preservatives (e.g., Thimerosal, benzyl alcohol, parabens),lubricants (e.g., stearic acid, magnesium stearate, polyethylene glycol,sodium lauryl sulfate), flow-aids (e.g., colloidal silicon dioxide),plasticizers (e.g., diethyl phthalate, triethyl citrate), emulsifiers(e.g., carbomer, hydroxypropyl cellulose, sodium lauryl sulfate),polymer coatings (e.g., poloxamers or poloxamines), coating and filmforming agents (e.g., ethyl cellulose, acrylates, polymethacrylates)and/or adjuvants.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

It is especially advantageous to formulate oral compositions in dosageunit form for ease of administration and uniformity of dosage. Dosageunit form as used herein refers to physically discrete units suited asunitary dosages for the subject to be treated; each unit containing apredetermined quantity of active compound calculated to produce thedesired therapeutic effect in association with the requiredpharmaceutical carrier. The specification for the dosage unit forms ofthe invention are dictated by and directly dependent on the uniquecharacteristics of the active compound and the particular therapeuticeffect to be achieved, and the limitations inherent in the art ofcompounding such an active compound for the treatment of individuals.

In one preferred embodiment, the compound can be formulated in anaqueous solution for intravenous injection. In one embodiment,solubilizing agents can be suitably employed. A particularly preferredsolubilizing agent includes cyclodextrins and modified cyclodextrins,such as sulfonic acid substituted β-cyclodextrin derivative or saltthereof.

In a particularly preferred embodiment, the inclusion complex comprisesa cyclodextrin and a compound selected from the group consisting ofcompound 12 and compound 18 or a geometric isomer, enantiomer,diastereomer, racemate, pharmaceutically acceptable salt, prodrug orsolvate thereof.

Cyclodextrins are cyclic oligomers of dextrose with a truncated conestructure consisting of a hydrophilic exterior and a hydrophobicinterior cavity. A cyclodextrin can form an inclusion complex with aguest molecule by complexing with all or a portion of a hydrophobicguest molecule within its cavity (for example, as described in, U.S.Pat. No. 4,727,064, the contents of which are herein incorporated byreference). The size of the cavity is determined by the number ofglucopyranose units in the cyclodextrin. Alpha- (α), beta- (B) andgamma- (γ) cyclodextrins are the most common cyclodextrins and possesssix, seven and eight glucopyranose units, respectively. Because naturalcyclodextrins have relatively low aqueous solubility and are associatedwith toxicity, chemically modified cyclodextrin derivatives have beendeveloped to overcome these limitations. Such cyclodextrin derivativestypically possess a chemical modification at one or more of the 2, 3, or6 position hydroxyl groups. Cyclodextrin derivatives have, for example,been described in U.S. Pat. Nos. 5,134,127, 5,376,645, 5,571,534,5,874,418, 6,046,177 and 6,133,248, the contents of which are hereinincorporated by reference and made a part hereof. As used herein, theterms “cyclodextrin,” “α-cyclodextrin,” “β-cyclodextrin and“γ-cyclodextrin” are intended to encompass unmodified cyclodextrins aswell as chemically modified derivatives thereof.

The compositions of the invention comprise an inclusion complex of acyclodextrin and a compound of Formulae (I), (II), (III) or (IV). In yetanother embodiment, the composition comprises a therapeuticallyeffective concentration of a compound of Formulae (I), (II), (III) or(IV). In a further embodiment, the composition further comprises apharmaceutically acceptable excipient or carrier.

In one embodiment of the invention, the composition comprises acyclodextrin selected from the group consisting of α-cyclodextrin,β-cyclodextrin and γ-cyclodextrin. In yet another embodiment, thecyclodextrin is a β-cyclodextrin and γ-cyclodextrin. In an additionalembodiment, the cyclodextrin is a β-cyclodextrin. In a furtherembodiment, the cyclodextrin is selected from the group consisting of a2-hydroxypropyl-β-cyclodextrin (Pitha et al, J Pharm Sci, 84 (8), 927-32(1995)) and sulfobutyl derivatized-β-cyclodextrin (described, forexample, in U.S. Pat. Nos. 5,134,127, 5,376,645, 5,874,418, 6,046,177and 6,133,248). In another embodiment, the cyclodextrin is a sulfobutylderivatized-β-cyclodextrin. One such sulfobutylderivatized-β-cyclodextrin is sulfobutylether-7-β-cyclodextrin and issold by CyDex, Inc. under the tradename CAPTISOL. In yet anotherembodiment of the invention, the cyclodextrin issulfobutylether-7-β-cyclodextrin.

The cyclodextrin may be included in an amount that increases thesolubility of the active compound in the composition. In one embodiment,the amount of cyclodextrin included within the composition is theminimal amount needed to solubilize the drug in the composition. In yetanother embodiment, the amount of cyclodextrin included within thecomposition is within about 5% of the minimal amount needed tosolubilize the drug. In a further embodiment, the composition is aparenteral formulation and the amount of cyclodextrin included withinthe formulation is the minimal amount of cyclodextrin needed tosolubilize the drug. In order to determine the minimum amount ofcyclodextrin needed to solubilize a compound encompassed by FormulaeI-IV, a plot of the compound's solubility versus cyclodextrinconcentration can be carried out. By interpolating or extrapolating fromthe plot, a composition can be prepared that contains the minimum amountof cyclodextrin needed to dissolve the desired concentration of theactive compound.

In one embodiment, the composition comprises at least about 0.5 or 1%(weight/volume) of a cyclodextrin. In another embodiment, thecomposition comprises at least about 5% of a cyclodextrin. In yetanother embodiment, the composition comprises at least about 15% of acyclodextrin. In a further embodiment, the composition comprises fromabout 0.5 to about 50% of a cyclodextrin. In yet another embodiment, thecomposition comprises from about 0.5% to about 40% of a cyclodextrin. Inanother embodiment, the composition comprises about 0.5% to about 35% ofa cycloextrin. In yet another embodiment, the composition comprisesabout 30% of a cyclodextrin.

In another embodiment, the composition comprises at least about 0.5 or1% (weight/volume) of a sulfobutyl derivatized-β-cyclodextrin. Inanother embodiment, the composition comprises at least about 5% of asulfobutyl derivatized-β-cyclodextrin. In yet another embodiment, thecomposition comprises at least about 15% of a sulfobutylderivatized-β-cyclodextrin. In a further embodiment, the compositioncomprises from about 0.5 to about 50% of a sulfobutylderivatized-β-cyclodextrin. In yet another embodiment, the compositioncomprises from about 0.5% to about 40% of a sulfobutylderivatized-β-cyclodextrin. In another embodiment, the compositioncomprises about 0.5% to about 35% of a sulfobutylderivatized-β-cyclodextrin. In yet another embodiment, the compositioncomprises about 30% of a sulfobutyl derivatized-β-cyclodextrin.

In one embodiment, the composition comprises at least about 0.5 or 1%(weight/volume) CAPTISOL. In another embodiment, the compositioncomprises at least about 5% CAPTISOL. In yet another embodiment, thecomposition comprises at least about 15% CAPTISOL. In a furtherembodiment, the composition comprises from about 0.5 to about 50%CAPTISOL. In yet another embodiment, the composition comprises fromabout 0.5% to about 40% CAPTISOL. In another embodiment, the compositioncomprises about 0.5% to about 35% CAPTISOL. In yet another embodiment,the composition comprises about 30% CAPTISOL.

In a further embodiment, the composition further comprises one or moreacids or bases. In one embodiment, the acid or base is added in anamount of 0.5 to 1.5 mol equivalents, preferably 1 to 1.3 molequivalents to formulate the compound. Acids that may be included in thecomposition include inorganic acids such as hydrochloric acid, sulfuricacid and phosphoric acid preferably hydrochloric acid, and organic acidssuch as citric acid, L(−)-malic acid and L(+)-tartaric acid preferablyL(+)-tartaric acid. Examples of bases that may be included in thecomposition include sodium hydroxide and potassium hydroxide preferablysodium hydroxide.

In a further embodiment, the composition optionally comprises dextran.In yet another embodiment, the composition comprises dextran in anamount of range from about 1% to about 5% weight/volume dextran. In afurther embodiment, the composition comprises from about 2 to about 4%weight/volume dextran.

The composition can be stored prior to administration to a patient. Inone embodiment, the composition is stored as a ready-to-use formulation.In yet another embodiment, the composition is stored having a dilutedconcentration of the active compound. The composition may be dilutedwith any appropriate excipient, for example, dextran or water. In afurther embodiment, the composition is stored having a higherconcentration of active compound for later dilution prior toadministration. An example of such a solubilizing agent is sold underthe trademark CAPTISOL® by CyDex, Inc. CAPTISOL is a polyanionicβ-cyclodextrin derivative with a sodium sulfonate salt separated fromthe lipophilic cavity by a butyl ether spacer group, or sulfobutylether(SBE) (as described in U.S. Pat. No. 5,134,127, the contents of whichare herein incorporated by reference). The selection of the SBE7-β-CD asthe cyclodextrin with the most desirable safety profile and drug carrierproperties was based upon evaluations of the mono, tetra andhepta-substituted preparations (SBE1, SBE4, and SBE7). CAPTISOL is thetrade name for CyDex's SBE7-β-CD PRODUCT.

Relative to β-cyclodextrin, the preferred solubilizing agents, such asCAPTISOL®, provide superior water solubility in excess of 70, preferably90 grams/100 ml.

In one embodiment, the solubilizing agent is added to the aqueoussolution in an amount of between about 0.5% and 50%, such as betweenabout 0.5% and 40% or about 0.5% and 35%, preferably between about 5%and 30%, more preferably between about 15 and 30%, such as about 30%weight/volume. Additional optional excipients can include dextran in anamount less than about 10%, such as less than about 5% weight/volume,such as about 5% weight/volume or between about 2 to 4% weight/volume.

Additional acids or bases may be added in an amount between about 0.5 to1.5 mol equivalents, preferably between about 1 to 1.3 mol equivalentsto further facilitate solubilization of the compound. Such acids caninclude inorganic acids such as hydrochloric acid, sulfuric acid andphosphoric acid preferably hydrochloric acid, and organic acids such ascitric acid, L(−)-malic acid and L(+)-tartaric acid preferablyL(+)-tartaric acid; such bases can include sodium hydroxide andpotassium hydroxide preferably sodium hydroxide.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

Daily administration may be repeated continuously for a period ofseveral days to several years. Oral treatment may continue for betweenone week and the life of the patient. Preferably the administration maytake place for five consecutive days after which time the patient can beevaluated to determine if further administration is required. Theadministration can be continuous or intermittent, e.g., treatment for anumber of consecutive days followed by a rest period. The compounds ofthe present invention may be administered intravenously on the first dayof treatment, with oral administration on the second day and allconsecutive days thereafter.

The preparation of pharmaceutical compositions that contain an activecomponent is well understood in the art, for example, by mixing,granulating, or tablet-forming processes. The active therapeuticingredient is often mixed with excipients that are pharmaceuticallyacceptable and compatible with the active ingredient. For oraladministration, the active agents are mixed with additives customary forthis purpose, such as vehicles, stabilizers, or inert diluents, andconverted by customary methods into suitable forms for administration,such as tablets, coated tablets, hard or soft gelatin capsules, aqueous,alcoholic or oily solutions and the like as detailed above.

The amount of the compound administered to the patient is less than anamount that would cause toxicity in the patient. In certain embodiments,the amount of the compound that is administered to the patient is lessthan the amount that causes a concentration of the compound in thepatient's plasma to equal or exceed the toxic level of the compound.Preferably, the concentration of the compound in the patient's plasma ismaintained at about 10 nM. In one embodiment, the concentration of thecompound in the patient's plasma is maintained at about 25 nM. In oneembodiment, the concentration of the compound in the patient's plasma ismaintained at about 50 nM. In one embodiment, the concentration of thecompound in the patient's plasma is maintained at about 100 nM. In oneembodiment, the concentration of the compound in the patient's plasma ismaintained at about 500 nM. In one embodiment, the concentration of thecompound in the patient's plasma is maintained at about 1000 nM. In oneembodiment, the concentration of the compound in the patient's plasma ismaintained at about 2500 nM. In one embodiment, the concentration of thecompound in the patient's plasma is maintained at about 5000 nM. Theoptimal amount of the compound that should be administered to thepatient in the practice of the present invention will depend on theparticular compound used and the type of cancer being treated.

DEFINITIONS

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “alkyl” embraces linear or branched radicals having one toabout twenty carbon atoms or, preferably, one to about twelve carbonatoms. More preferred alkyl radicals are “lower alkyl” radicals havingone to about ten carbon atoms. Most preferred are lower alkyl radicalshaving one to about eight carbon atoms. Examples of such radicalsinclude methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl and the like.

The term “alkenyl” embraces linear or branched radicals having at leastone carbon-carbon double bond of two to about twenty carbon atoms or,preferably, two to about twelve carbon atoms. More preferred alkenylradicals are “lower alkenyl” radicals having two to about ten and morepreferably about two to about eight carbon atoms. Examples of alkenylradicals include ethenyl, allyl, propenyl, butenyl and 4-methylbutenyl.The terms “alkenyl”, and “lower alkenyl”, embrace radicals having “cis”and “trans” orientations, or alternatively, “E” and “Z” orientations.

The term “alkynyl” embraces linear or branched radicals having at leastone carbon-carbon triple bond of two to about twenty carbon atoms or,preferably, two to about twelve carbon atoms. More preferred alkynylradicals are “lower alkynyl” radicals having two to about ten and morepreferably two to about eight carbon atoms. Examples of alkynyl radicalsinclude propargyl, 1-propynyl, 2-propynyl, 1-butyne, 2-butynyl and1-pentynyl.

The term “alkoxy” embraces linear or branched oxy-containing radicalseach having alkyl portions of one to about twenty carbon atoms or,preferably, one to about twelve carbon atoms. More preferred alkoxyradicals are “lower alkoxy” radicals having one to about ten and morepreferably having one to about eight carbon atoms. Examples of suchradicals include methoxy, ethoxy, propoxy, butoxy and tert-butoxy.

The term “alkoxyalkoxy” embraces alkoxy radicals having one or morealkoxy radicals attached to the alkyl radical, that is, to formmonoalkoxyalkyl and dialkoxyalkyl radicals.

The term “substituted” refers to the replacement of one or more hydrogenradicals in a given structure with the radical of a specifiedsubstituent including, but not limited to: halo, alkyl, alkenyl,alkynyl, aryl, heterocyclyl, thiol, alkylthio, arylthio, alkylthioalkyl,arylthioalkyl, alkylsulfonyl, alkylsulfonylalkyl, arylsulfonylalkyl,alkoxy, aryloxy, aralkoxy, aminocarbonyl, alkylaminocarbonyl,arylaminocarbonyl, alkoxycarbonyl, aryloxycarbonyl, haloalkyl, amino,trifluoromethyl, cyano, nitro, alkylamino, arylamino, alkylaminoalkyl,arylaminoalkyl, aminoalkylamino, hydroxy, alkoxyalkyl, carboxyalkyl,alkoxycarbonylalkyl, aminocarbonylalkyl, acyl, aralkoxycarbonyl,carboxylic acid, sulfonic acid, sulfonyl, phosphonic acid, aryl,heteroaryl, heterocyclic, and aliphatic. It is understood that thesubstituent can be further substituted.

The terms “halogen” or “halo” as used herein, refers to an atom selectedfrom fluorine, chlorine, bromine and iodine.

As used herein, the term “aberrant proliferation” refers to abnormalcell growth.

The phrase “adjunctive therapy” encompasses treatment of a subject withagents that reduce or avoid side effects associated with the combinationtherapy of the present invention, including, but not limited to, thoseagents, for example, that reduce the toxic effect of anticancer drugs,e.g., bone resorption inhibitors, cardioprotective agents; prevent orreduce the incidence of nausea and vomiting associated withchemotherapy, radiotherapy or operation; or reduce the incidence ofinfection associated with the administration of myelosuppressiveanticancer drugs.

The term “angiogenesis,” as used herein, refers to the formation ofblood vessels. Specifically, angiogenesis is a multi-step process inwhich endothelial cells focally degrade and invade through their ownbasement membrane, migrate through interstitial stroma toward anangiogenic stimulus, proliferate proximal to the migrating tip, organizeinto blood vessels, and reattach to newly synthesized basement membrane(see Folkman et al., Adv. Cancer Res., Vol. 43, pp. 175-203 (1985)).Anti-angiogenic agents interfere with this process. Examples of agentsthat interfere with several of these steps include thrombospondin-1,angiostatin, endostatin, interferon alpha and compounds such as matrixmetalloproteinase (MMP) inhibitors that block the actions of enzymesthat clear and create paths for newly forming blood vessels to follow;compounds, such as .alpha.v.beta.3 inhibitors, that interfere withmolecules that blood vessel cells use to bridge between a parent bloodvessel and a tumor; agents, such as specific COX-2 inhibitors, thatprevent the growth of cells that form new blood vessels; andprotein-based compounds that simultaneously interfere with several ofthese targets.

The term “apoptosis” as used herein refers to programmed cell death assignaled by the nuclei in normally functioning human and animal cellswhen age or state of cell health and condition dictates. An “apoptosisinducing agent” triggers the process of programmed cell death.

The term “cancer” as used herein denotes a class of diseases ordisorders characterized by uncontrolled division of cells and theability of these cells to invade other tissues, either by direct growthinto adjacent tissue through invasion or by implantation into distantsites by metastasis.

The term “compound” is defined herein to include pharmaceuticallyacceptable salts, solvates, hydrates, polymorphs, enantiomers,diastereoisomers, racemates and the like of the compounds having aformula as set forth herein.

The term “devices” refers to any appliance, usually mechanical orelectrical, designed to perform a particular function.

As used herein, the term “dysplasia” refers to abnormal cell growth andtypically refers to the earliest form of pre-cancerous lesionrecognizable in a biopsy by a pathologist.

As used herein, the term “effective amount of the subject compounds,”with respect to the subject method of treatment, refers to an amount ofthe subject compound which, when delivered as part of desired doseregimen, brings about, e.g. a change in the rate of cell proliferationand/or state of differentiation and/or rate of survival of a cell toclinically acceptable standards. This amount may further relieve to someextent one or more of the symptoms of a neoplasia disorder, including,but is not limited to: 1) reduction in the number of cancer cells; 2)reduction in tumor size; 3) inhibition (i.e., slowing to some extent,preferably stopping) of cancer cell infiltration into peripheral organs;4) inhibition (i.e., slowing to some extent, preferably stopping) oftumor metastasis; 5) inhibition, to some extent, of tumor growth; 6)relieving or reducing to some extent one or more of the symptomsassociated with the disorder; and/or 7) relieving or reducing the sideeffects associated with the administration of anticancer agents.

The term “hyperplasia,” as used herein, refers to excessive celldivision or growth.

The phrase an “immunotherapeutic agent” refers to agents used totransfer the immunity of an immune donor, e.g., another person or ananimal, to a host by inoculation. The term embraces the use of serum orgamma globulin containing performed antibodies produced by anotherindividual or an animal; nonspecific systemic stimulation; adjuvants;active specific immunotherapy; and adoptive immunotherapy. Adoptiveimmunotherapy refers to the treatment of a disease by therapy or agentsthat include host inoculation of sensitized lymphocytes, transferfactor, immune RNA, or antibodies in serum or gamma globulin.

The term “inhibition,” in the context of neoplasia, tumor growth ortumor cell growth, may be assessed by delayed appearance of primary orsecondary tumors, slowed development of primary or secondary tumors,decreased occurrence of primary or secondary tumors, slowed or decreasedseverity of secondary effects of disease, arrested tumor growth andregression of tumors, among others. In the extreme, complete inhibition,is referred to herein as prevention or chemoprevention.

The term “metastasis,” as used herein, refers to the migration of cancercells from the original tumor site through the blood and lymph vesselsto produce cancers in other tissues. Metastasis also is the term usedfor a secondary cancer growing at a distant site.

The term “neoplasm,” as used herein, refers to an abnormal mass oftissue that results from excessive cell division. Neoplasms may bebenign (not cancerous), or malignant (cancerous) and may also be calleda tumor. The term “neoplasia” is the pathological process that resultsin tumor formation.

As used herein, the term “pre-cancerous” refers to a condition that isnot malignant, but is likely to become malignant if left untreated.

The term “proliferation” refers to cells undergoing mitosis.

The phrase “EGFR-TK related disease or disorder” refers to a disease ordisorder characterized by inappropriate EGFR-TK activity orover-activity of the EGFR-TK. Inappropriate activity refers to either;(i) EGFR-TK expression in cells which normally do not express EGFR-TKs;(ii) increased EGFR-TK expression leading to unwanted cellproliferation, differentiation and/or growth; or, (iii) decreasedEGFR-TK expression leading to unwanted reductions in cell proliferation,differentiation and/or growth. Over-activity of EGFR-TKs refers toeither amplification of the gene encoding a particular EGFR-TK orproduction of a level of EGFR-TK activity which can correlate with acell proliferation, differentiation and/or growth disorder (that is, asthe level of the EGFR-TK increases, the severity of one or more of thesymptoms of the cellular disorder increases). Over activity can also bethe result of ligand independent or constitutive activation as a resultof mutations such as deletions of a fragment of a EGFR-TK responsiblefor ligand binding.

The phrase a “radio therapeutic agent” refers to the use ofelectromagnetic or particulate radiation in the treatment of neoplasia.

The term “recurrence” as used herein refers to the return of cancerafter a period of remission. This may be due to incomplete removal ofcells from the initial cancer and may occur locally (the same site ofinitial cancer), regionally (in vicinity of initial cancer, possibly inthe lymph nodes or tissue), and/or distally as a result of metastasis.

The term “treatment” refers to any process, action, application,therapy, or the like, wherein a mammal, including a human being, issubject to medical aid with the object of improving the mammal'scondition, directly or indirectly.

The term “vaccine” includes agents that induce the patient's immunesystem to mount an immune response against the tumor by attacking cellsthat express tumor associated antigens (Teas).

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid or inorganic acid. Examples of pharmaceuticallyacceptable nontoxic acid addition salts include, but are not limited to,salts of an amino group formed with inorganic acids such as hydrochloricacid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloricacid or with organic acids such as acetic acid, maleic acid, tartaricacid, citric acid, succinic acid lactobionic acid or malonic acid or byusing other methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include, but are not limited to,adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Representative alkali or alkaline earth metal salts includesodium, lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include, but are not limited to, formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds of the present invention which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals with undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, and effective for their intended use, aswell as the zwitterionic forms, where possible, of the compounds of thepresent invention. “Prodrug”, as used herein means a compound which isconvertible in vivo by metabolic means (e.g. by hydrolysis) to acompound of the invention. Various forms of prodrugs are known in theart, for example, as discussed in Bundgaard, (ed.), Design of Prodrugs,Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, vol. 4,Academic Press (1985); Krogsgaard-Larsen, et al., (ed). “Design andApplication of Prodrugs, Textbook of Drug Design and Development,Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug DeliverReviews, 8:1-38 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel DrugDelivery Systems, American Chemical Society (1975); and Bernard Testa &Joachim Mayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry,Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

As used herein, “pharmaceutically acceptable carrier” is intended toinclude any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents, and thelike, compatible with pharmaceutical administration, such as sterilepyrogen-free water. Suitable carriers are described in the most recentedition of Remington's Pharmaceutical Sciences, a standard referencetext in the field, which is incorporated herein by reference. Preferredexamples of such carriers or diluents include, but are not limited to,water, saline, finger's solutions, dextrose solution, and 5% human serumalbumin. Liposomes and non-aqueous vehicles such as fixed oils may alsobe used. The use of such media and agents for pharmaceutically activesubstances is well known in the art. Except insofar as any conventionalmedia or agent is incompatible with the active compound, use thereof inthe compositions is contemplated. Supplementary active compounds canalso be incorporated into the compositions.

As used herein, the term “pre-cancerous” refers to a condition that isnot malignant, but is likely to become malignant if left untreated.

The term “subject” as used herein refers to an animal. Preferably theanimal is a mammal. More preferably the mammal is a human. A subjectalso refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and may include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the formulae herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons(1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The compounds described herein may contain one or more asymmetriccenters and thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds, otherunsaturation, or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers and/or cis- and trans-isomers. Likewise, alltautomeric forms are also intended to be included. The configuration ofany carbon-carbon double bond appearing herein is selected forconvenience only and is not intended to designate a particularconfiguration unless the text so states; thus a carbon-carbon doublebond or carbon-heteroatom double bond depicted arbitrarily herein astrans may be cis, trans, or a mixture of the two in any proportion.

In some embodiments, the inventive compound is a crystalline or anamorphous form of a compound of the invention. In another embodiment,the inventive compound is a mixture of two or more crystalline forms ofthe compound. In a further embodiment, the invention is a crystallineform or a mixture of two or more crystalline forms of compound 12. Acrystalline form of compound 12 can be prepared substantially asdescribed in Example 8 (Method 2) below.

Different crystalline forms of chemical compounds (or polymorphs) eachexhibit different physical, chemical, spectroscopic and/orcrystallographic properties. In one embodiment, the crystalline form ofthe invention has an X-ray powder diffraction (XRPD) pattern with atleast three, four, five, six, seven or eight major peaks in common withthe X-ray powder diffraction pattern of the crystal prepared accordingto working Example 8 (Method 2). A major peak is an XRPD peak with arelative intensity greater than about 25%; relative intensity iscalculated as a ratio of the peak intensity of the peak of interestversus the peak intensity of the largest peak. As used herein, a peakfor the crystalline form is in common with the XRPD pattern of thecrystal prepared according to Example 8 (Method 2) if it is within 0.5°20 of a peak location for the crystal prepared according to Example 8(Method 2). In another embodiment, the crystalline form of compound 12has an endothermic transition within about 1.0° C. as the crystallineform prepared according to Example 8 (Method 2). In yet anotherembodiment, the crystalline form of compound 12 has an endothermictransition (as observed by differential scanning calorimetry) withinabout 0.5° C. of as the crystal formed according to Example 8 (Method 2)and at least three, four, five, six, seven or eight major peaks incommon with the XRPD pattern of the compound prepared according toExample 8 (Method 2). In a further embodiment, the crystalline form ofcompound 12 exhibits a differential scanning calorimetry pattern that issubstantially the same as the differential scanning calorimetry patternof the compound produced according the Example 8 (Method 2).

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers or excipients.

As used herein, the term “pharmaceutically acceptable carrier orexcipient” means a non-toxic, inert solid, semi-solid or liquid filler,diluent, encapsulating material or formulation auxiliary of any type.Some examples of materials which can serve as pharmaceuticallyacceptable carriers are sugars such as lactose, glucose and sucrose;starches such as corn starch and potato starch; cellulose and itsderivatives such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; powdered tragacanth; malt; gelatin; talc; excipientssuch as cocoa butter and suppository waxes; oils such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; glycols such as propylene glycol; esters such as ethyloleate and ethyl laurate; agar; buffering agents such as magnesiumhydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffersolutions, as well as other non-toxic compatible lubricants such assodium lauryl sulfate and magnesium stearate, as well as solubilizingagents, coloring agents, releasing agents, coating agents, sweetening,flavoring and perfuming agents, preservatives and antioxidants can alsobe present in the composition, according to the judgment of theformulator.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions, may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

For pulmonary delivery, a therapeutic composition of the invention isformulated and administered to the patient in solid or liquidparticulate form by direct administration e.g., inhalation into therespiratory system. Solid or liquid particulate forms of the activecompound prepared for practicing the present invention include particlesof respirable size: that is, particles of a size sufficiently small topass through the mouth and larynx upon inhalation and into the bronchiand alveoli of the lungs. Delivery of aerosolized therapeutics,particularly aerosolized antibiotics, is known in the art (see, forexample U.S. Pat. No. 5,767,068 to VanDevanter et al., U.S. Pat. No.5,508,269 to Smith et al., and WO 98/43,650 by Montgomery, all of whichare incorporated herein by reference). A discussion of pulmonarydelivery of antibiotics is also found in U.S. Pat. No. 6,014,969,incorporated herein by reference.

By a “therapeutically effective amount” of a compound of the inventionis meant an amount of the compound which confers a therapeutic effect onthe treated subject, at a reasonable benefit/risk ratio applicable toany medical treatment. The therapeutic effect may be objective (i.e.,measurable by some test or marker) or subjective (i.e., subject gives anindication of or feels an effect). An effective amount of the compounddescribed above may range from about 0.1 mg/Kg to about 500 mg/Kg,preferably from about 1 to about 50 mg/Kg. Effective doses will alsovary depending on route of administration, as well as the possibility ofco-usage with other agents. It will be understood, however, that thetotal daily usage of the compounds and compositions of the presentinvention will be decided by the attending physician within the scope ofsound medical judgment. The specific therapeutically effective doselevel for any particular patient will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the activity of the specific compound employed; the specific compositionemployed; the age, body weight, general health, sex and diet of thepatient; the time of administration, route of administration, and rateof excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or contemporaneously with thespecific compound employed; and like factors well known in the medicalarts.

The total daily dose of the compounds of this invention administered toa human or other animal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

The compounds of the formulae described herein can, for example, beadministered by injection, intravenously, intraarterially, subdermally,intraperitoneally, intramuscularly, or subcutaneously; or orally,buccally, nasally, transmucosally, topically, in an ophthalmicpreparation, or by inhalation, with a dosage ranging from about 0.1 toabout 500 mg/kg of body weight, alternatively dosages between 1 mg and1000 mg/dose, every 4 to 120 hours, or according to the requirements ofthe particular drug. The methods herein contemplate administration of aneffective amount of compound or compound composition to achieve thedesired or stated effect. Typically, the pharmaceutical compositions ofthis invention will be administered from about 1 to about 6 times perday or alternatively, as a continuous infusion. Such administration canbe used as a chronic or acute therapy. The amount of active ingredientthat may be combined with pharmaceutically excipients or carriers toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. A typical preparation willcontain from about 5% to about 95% active compound (w/w). Alternatively,such preparations may contain from about 20% to about 80% activecompound.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Patients may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

Synthetic Methods

A quinazoline derivative of the formula I, or apharmaceutically-acceptable salt thereof, may be prepared by any processknown to be applicable to the preparation of chemically-relatedcompounds. Suitable processes for making certain intermediates include,for example, those illustrated in European Patent Applications Nos.0520722, 0566226, 0602851, 0635498, 0635507, U.S. Pat. Nos. 5,457,105,5,770,599, US publication No. 2003/0158408 and reference such as, J. MedChem. 2004, 47, 871-887. Necessary starting materials may be obtained bystandard procedures of organic chemistry. The preparation of suchstarting materials is described within the accompanying non-limitingExamples. Alternatively necessary starting materials are obtainable byanalogous procedures to those illustrated which are within the ordinaryskill of a chemist.

The compounds and processes of the present invention will be betterunderstood in connection with the following representative syntheticschemes that illustrate the methods by which the compounds of theinvention may be prepared, which are intended as an illustration onlyand not limiting of the scope of the invention.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Example 1 Preparation of2-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyyquinazolin-6-yloxy)-N-hydroxyacetamide(Compound 1) Step 1a. 6,7-Dimethoxyquinazolin-4(3H)-one (Compound 0102)

A mixture of methyl 2-amino-4,5-dimethoxybenzoic acid 0101 (2.1 g, 10mmol), ammonium formate (0.63 g, 10 mmol) and formamide (7 ml) wasstirred and heated to 190˜200° C. for 2 hours. Then the mixture wascooled to room temperature. The precipitate was isolated, washed withwater and dried to provide the title compound 0102 as a brown solid (1.8g, 84.7%): LCMS: m/z 207 [M+1]⁺; ¹H NMR (DMSO) δ 3.87 (s, 3H), 3.89 (s,3H), 7.12 (s, 1H), 7.43 (s, 1H), 7.97 (s, 1H), 12.08 (bs, 1H).

Step 1b. 6-Hydroxy-7-methoxyquinazolin-4(3H)-one (Compound 0103)

6,7-Dimethoxyquinazolin-4(3H)-one (0102) (10.3 g, 50 mmol) was addedportionwise to stirred methanesulphonic acid (68 ml). L-Methionone (8.6g, 57.5 mmol) was then added and resultant mixture was heated to150˜160° C. for 5 hours. The mixture was cooled to room temperature andpoured onto a mixture (250 ml) of ice and water. The mixture wasneutralized by the addition of aqueous sodium hydroxide solution (40%).The precipitate was isolated, washed with water and dried to yield titlecompound 0103 as a grey solid (10 g, crude): LCMS: m/z 193 [M+1]⁺.

Step 1c. 3,4-Dihydro-7-methoxy-4-oxoquinazolin-6-yl acetate (Compound0104)

A mixture of 6-hydroxy-7-methoxyquinazolin-4(3H)-one (0103) (10 gcrude), acetic anhydride (100 ml) and pyridine (8 ml) was stirred andheated to reflux for 3 hours. The mixture was cooled to room temperatureand poured into a mixture (250 ml) of ice and water. The precipitate wasisolated and dried to yield the title product 0104 as a grey solid (5.8g, 50% two step overall yield): LCMS: m/z 235 [M+1]⁺; ¹H NMR (CDCl₃) δ2.27 (s, 3H), 3.89 (s, 3H), 7.28 (s, 1H), 7.72 (s, 1H), 8.08 (d, 1H),12.20 (bs, 1H).

Step 1d. 4-Chloro-7-methoxyquinazolin-6-yl acetate (Compound 0105)

A mixture of 3,4-dihydro-7-methoxy-4-oxoquinazolin-6-yl acetate (0104)(2.0 g, 8.5 mmol) and phosphoryl trichloride (20 ml) was stirred andheated to reflux for 3 hours. When a clear solution was obtained, theexcessive phosphoryl trichloride was removed under reduced pressure. Theresidue was dissolved in dichloromethane (50 ml) and the organic layerwas washed with aqueous NaHCO₃ solution (20 ml×2) and brine (20 ml×1)and dried over MgSO₄, filtered and evaporated to give the title product0105 as a yellow solid (1.4 g, 65%): LCMS: m/z 249 [M+1]⁺; ¹H NMR(CDCl₃) δ 2.40 (s, 3H), 4.03 (s, 3H), 7.44 (s, 1H), 7.90 (s, 1H), 8.95(bs, 1H).

Step 1e. 4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-ylacetate hydrochloride (Compound 0108)

A mixture of 4-chloro-7-methoxyquinazolin-6-yl acetate (0105) (1.3 g,5.1 mmol) and 3-chloro-4-fluorobenzenamine 0106 (1.5 g, 10.2 mmol) inisopropanol (45 ml) was stirred and heated to reflux for 3 hours. Themixture was cooled to room temperature and resulting precipitate wasisolated. The solid was then dried to give the title compound 0108 as alight yellow solid (1.6 g, 79%): LCMS: m/z 362 [M+1]⁺; ¹H NMR (DMSO) δ2.36 (s, 3H), 3.98 (s, 3H), 7.49 (s, 1H), 7.52 (d, 1H), 7.72 (m, 1H),8.02 (dd, 1H), 8.71 (s, 1H), 8.91 (s, 1H), 11.4 (bs, 1H).

Step 1f. 4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-ol(Compound 0109)

A mixture of compound (0107) (1.41 g, 3.5 mmol), LiOH H₂O (0.5 g, 11.7mmol) in methanol (100 ml) and H₂O (100 ml) was stirred at roomtemperature for 0.5 hour. The mixture was neutralized by addition ofdilution acetic acid. The precipitate was isolated and dried to give thetitle compound 0109 as a grey solid (1.06 g, 94%): LCMS: m/z 320 [M+1]⁺;¹H NMR (DMSO) δ 3.99 (s, 3H), 7.20 (s, 1H), 7.38 (t, 1H), 7.75 (s, 1H),7.81 (m, 1H), 8.20 (m, 1H), 8.46 (s, 1H), 9.46 (s, 1H), 9.68 (s, 1H).

Step 1g. Ethyl2-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)acetate(Compound 0110-1)

A mixture of compound 0109 (300 mg, 0.94 mmol) and Ethyl 2-bromoacetate(163 mg, 0.98 mmol) and potassium carbonate (323 mg, 2.35 mmol) inN,N-dimethylformamide (6 ml) was stirred and heated to 40° for 30minutes. The reaction process was monitored by TLC. The mixture wasfiltrated. The filtration was concentrated under reduce pressure. Theresidues was wash with diethyl ether and dried to give the titlecompound 0110-1 as a yellow solid (280 mg, 74%): LCMS: m/z 406 [M+1]⁺;¹H NMR (DMSO) δ 1.23 (t, 3H), 3.96 (s, 3H), 4.20 (q, 2H), 4.95 (s, 2H),7.24 (s, 1H), 7.44 (t, 1H), 7.75 (m, 1H), 7.82 (s, 1H), 8.10 (dd, 1H),8.51 (s, 1H), 9.54 (s, 1H).

Step 1 h.2-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyacetamide(Compound 1)

To a stirred solution of hydroxyamine hydrochloride (4.67 g, 67 mmol) inmethanol (24 ml) at 0° C. was added a solution of potassium hydroxide(5.61 g, 100 mmol) in methanol (14 ml). After addition, the mixture wasstirred for 30 minutes at 0° C., and was allowed to stand at lowtemperature. The resulting precipitate was isolated, and the solutionwas prepared to give free hydroxyamine.

The above freshly prepared hydroxyamine solution (1.4 ml, 2.4 mmol) wasplaced in 5 ml flask. Compound 0110-1 (250 mg, 0.6 mmol) was added tothis solution and stirred at 0° C. for 10 minutes, and raise to roomtemperature. The reaction process was monitored by TLC. The mixture wasneutralized with acetic acid. The mixture was concentrated under reducepressure. The residue was purified by preparation HPLC. To give thetitle compound 1 as a grey solid (50 mg, 21%): LCMS: m/z 393 [M+1]⁺; ¹HNMR (DMSO) δ 3.96 (s, 3H), 4.62 (s, 2H), 7.24 (s, 1H), 7.45 (t, 1H),7.78 (m, 1H), 7.86 (s, 1H), 8.10 (dd, 1H), 8.52 (s, 1H), 9.07 (s, 1H),9.57 (s, 1H), 10.80 (s, 1H).

Example 2 Preparation of4-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxybutanamide(Compound 3) Step 2a. Ethyl4-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)butanoate(Compound 0110-3)

The title compound 0110-3 was prepared as a yellow solid (220 mg, 80.5%)from compound 0109 from step 1f (200 mg, 0.63 mmol) and ethyl4-bromobutyrate (135 mg, 0.69 mmol) using a procedure similar to thatdescribed for compound 0110-1 (example 1): LCMS: m/z 434 [M+1]⁺; ¹H NMR(CDCl₃) δ 1.36 (t, 3H), 2.23 (m, 2H), 2.57 (t, 2H), 4.03 (s, 3H), 4.32(m, 4H), 7.15 (t, 1H), 7.25 (m, 1H), 7.87 (s, 1H), 8.00 (m, 2H), 8.15(bs, 1H), 8.57 (s, 1H).

Step 2b.4-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxybutanamide(Compound 3)

The title compound 3 was prepare as a grey solid (25 mg, 12%) fromcompound 0110-3 (200 mg, 0.23 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: m/z 421 [M+1]⁺; ¹H NMR(DMSO): δ 2.06 (m, 2H), 2.22 (t, 2H), 3.95 (s, 3H), 4.15 (t, 2H), 7.21(s, 1H), 7.43 (t, 1H), 7.83 (s, 2H), 8.14 (dd, 1H), 8.51 (s, 1H), 8.75(s, 1H), 9.56 (s, 1H), 10.50 (s, 1H).

Example 3 Preparation of7-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyhexanamide(Compound 5) Step 3a. Ethyl6-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)hexanoate(Compound 0110-5)

The title compound 0110-5 was prepared as a yellow solid (510 mg, 68%)from compound 0109 from step 1f (510 mg, 1.6 mmol) and ethyl6-bromohexanoate (430 mg, 1.9 mmol) using a procedure similar to thatdescribed for compound 0110-1 (Example 1): LCMS: m/z 462 [M+1]⁺; ¹H NMR(CDCl₃): δ 1.24 (t, 3H), 1.55 (m, 2H), 1.74 (m, 2H), 1.91 (m, 2H), 2.38(m, 2H), 3.97 (s, 3H), 4.13 (m, 4H), 7.15 (t, 1H), 7.25 (m, 2H), 7.60(m, 1H), 7.86 (m, 1H), 7.91 (dd, 1H), 8.61 (s, 1H).

Step 3b.7-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyhexanamide(Compound 5)

The title compound 5 was prepared as a grey solid (100 mg, 34%) formcompound 0110-5 (305 mg, 0.66 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): m.p. 206.6˜207.1° C. (dec); LCMS:m/z 449 [M+1]⁺; ¹H NMR (DMSO) δ 1.44 (m, 2H), 1.64 (m, 2H), 1.82 (m,2H), 1.99 (t, 2H), 3.93 (s, 3H), 4.12 (t, 2H), 7.19 (s, 1H), 7.43 (t,1H), 7.79 (m, 2H), 8.12 (dd, 1H), 8.49 (s, 1H), 8.68 (s, 1H), 9.53 (s,1H), 10.37 (s, 1H).

Example 4 Preparation of7-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 6) Step 4a. Ethyl7-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(compound 0110-6)

The title compound 0110-6 was prepared as a yellow solid (390 mg, 53%)from compound 0109 from step 1f (512 mg, 1.6 mmol) and ethyl7-bromoheptanoate (438 mg, 1.8 mmol) using a procedure similar to thatdescribed for compound 0110-1 (Example 1): LCMS: m/z 476 [M+1]⁺; ¹H NMR(CDCl₃) δ 1.24 (t, 3H), 1.43 (m, 4H), 1.66 (m, 2H), 1.88 (m, 2H), 2.32(t, 2H), 3.97 (s, 3H), 4.07 (t, 2H), 4.12 (q, 2H), 7.15 (t, 1H), 7.23(t, 2H), 7.66 (m, 1H), 7.75 (m, 1H), 7.87 (dd, 1H), 8.65 (s, 1H).

Step 4b.7-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(compound 6)

The title compound 6 was prepared as a grey solid (80 mg, 25%) fromcompound 0110-6 (323 mg, 0.68 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): m.p. 180.8˜182.3° C. (dec); LCMS:m/z 463 [M+1]⁺; ¹H NMR (DMSO) δ 1.34 (m, 2H), 1.50 (m, 4H), 1.81 (m,2H), 1.96 (t, 2H), 3.92 (s, 3H), 4.11 (t, 2H), 7.18 (s, 1H), 7.43 (t,1H), 7.78 (m, 2H), 8.12 (dd, 1H), 8.48 (s, 1H), 8.64 (s, 1H), 9.50 (s,1H), 10.33 (s, 1H).

Example 5 Preparation of2-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyacetamide(Compound 7) Step 5a. 4-(3-Ethynylphenylamino)-7-methoxyquinazolin-6-ylacetate Hydrochloride (Compound 0111)

A mixture of 4-chloro-7-methoxyquinazolin-6-yl acetate (0105) (2.6 g,10.2 mmol) and 3-ethynylbenzenamine (0107) (2.4 g, 20.5 mmol) inisopropanol (100 ml) was stirred and heated to reflux for 3 hours. Themixture was cooled to room temperature. The precipitate was isolated anddried to give the title compound 0111 as a yellow solid (2.6 g, 68%):LCMS: m/z 334 [M+1]⁺; ¹H NMR (DMSO) δ 2.39 (s, 3H), 3.17 (s, 1H), 3.98(s, 3H), 7.35 (m, 1H), 7.40 (s, 1H), 7.47 (m, 1H), 7.72 (m, 1H), 7.90(s, 1H), 8.57 (s, 1H), 8.87 (s, 1H), 10.99 (bs, 1H).

Step 5b. 4-(3-Ethynylphenylamino)-7-methoxyquinazolin-6-ol (Compound0112)

A mixture of compound 0111 (2.0 g, 5.4 mmol) and LiOH H₂O (0.75 g, 17.9mmol) in methanol (100 ml) and H₂O (100 ml) was stirred at roomtemperature for 0.5 hour. The mixture was neutralized by addition ofdilution acetic acid. The precipitate was isolated and dried to give thetitle compound 0112 as a grey solid (1.52 g, 96%): LCMS: m/z 292 [M+1]⁺;¹H NMR (DMSO) δ 3.17 (s, 1H), 3.98 (s, 3H), 7.18 (d, 1H), 7.21 (s, 1H),7.37 (t, 1H), 7.80 (s, 1H), 7.90 (d, 1H), 8.04 (m, 1H), 8.47 (s, 1H),9.41 (s, 1H), 9.68 (bs, 1H).

Step 5c. Ethyl2-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)acetate(Compound 0113-7)

The title compound 0113-7 was prepared as a yellow solid (450 mg, 69%)from compound 0112 (500 mg, 1.72 mmol) and ethyl 2-bromoacetate (300 mg,1.8 mmol) using a procedure similar to that described for compound0110-1 (Example 1): LCMS: m/z 378 [M+1]⁺; ¹H NMR (DMSO) δ 1.22 (t, 3H),3.97 (s, 3H), 4.21 (q, 2H), 4.97 (t, 2H), 7.22 (d, 1H), 7.24 (s, 1H),7.42 (t, 1H), 7.84 (m, 2H), 7.86 (d, 1H), 7.96 (s, 1H), 8.51 (s, 1H).

Step 5d.2-(4-(3-Ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyacetamide(Compound 7)

The title compound 7 was prepared as a grey solid (100 mg, 23%) fromcompound 0113-7 (448 mg, 1.2 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: m/z 365 [M+1]⁺; ¹H NMR(DMSO) δ 4.00 (s, 3H), 4.26 (s, 1H), 4.65 (s, 2H), 7.27 (s, 1H), 7.37(d, 1H), 7.49 (t, 1H), 7.73 (d, 1H), 7.85 (s, 1H), 8.03 (s, 1H), 8.78(s, 1H), 9.17 (bs, 1H), 10.60 (s, 1H), 10.85 (s, 1H).

Example 6 Preparation of4-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxybutanamide(Compound 9) Step 6a. Ethyl4-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)butanoate(Compound 0113-9)

The title compound 0113-9 was prepared as a yellow solid (438 mg, 59%)from compound 0112 (500 mg, 1.72 mmol) and ethyl 4-bromobutyrate (349mg, 1.8 mmol) using a procedure similar to that described for compound0110-1 (Example 1): LCMS: m/z 406 [M+1]⁺; ¹H NMR (CDCl₃) δ 1.37 (t, 3H),2.34 (m, 2H), 2.56 (t, 2H), 3.07 (s, 1H), 4.03 (s, 3H), 4.32 (m, 4H),7.21 (m, 1H), 7.25 (s, 1H), 7.36 (t, 1H), 7.94 (s, 1H), 7.97 (m, 1H),8.20 (s, 1H), 8.28 (m, 1H), 8.70 (s, 1H).

Step 6b.4-(4-(3-Ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxybutanamide(Compound 9)

The title compound 9 was prepared as a grey solid (60 mg, 31%) fromcompound 0113-9 (200 mg, 0.49 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: m/z 393 [M+1]⁺; ¹H NMR(DMSO) δ 2.06 (m, 2H), 2.22 (t, 2H), 3.30 (s, 1H), 3.95 (s, 3H), 4.16(t, 2H), 7.19 (m, 2H), 7.40 (t, 1H), 7.85 (s, 1H), 7.91 (d, 1H), 8.02(s, 1H), 8.51 (s, 1H), 8.74 (s, 1H), 9.49 (s, 1H), 10.49 (s, 1H).

Example 7 Preparation of6-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyhexanamide(Compound 11) Step 7a. Ethyl6-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)hexanoate(Compound 0113-11)

The title compound 0113-11 was prepared as yellow solid (543 mg, 73%)from compound 0112 from step 5b (500 mg, 1.72 mmol) and ethyl6-bromohexanoate (401 mg, 1.8 mmol) using a procedure similar to thatdescribed for compound 0110-1 (Example 1): LCMS: m/z 434 [M+1]⁺; ¹H NMR(CDCl₃) δ 1.24 (t, 3H), 1.53 (m, 2H), 1.72 (m, 2H), 1.90 (m, 2H), 2.37(t, 3H), 3.08 (s, 1H), 3.97 (s, 3H), 4.10 (m, 4H), 7.19 (s, 1H), 7.25(m, 2H), 7.34 (t, 1H), 7.67 (s, 1H), 7.78 (m, 1H), 7.84 (m, 1H), 8.67(s, 1H).

Step 7b.6-(4-(3-Ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyhexanamide(Compound 11)

The title compound 11 was prepared as a grey solid (110 mg, 41%) fromcompound 0113-11 (275 mg, 0.63 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): m.p. 193.4˜195.8° C. (dec); LCMS:m/z 421 [M+1]⁺; ¹H NMR (DMSO) δ 1.44 (m, 2H), 1.60 (m, 2H), 1.84 (m,2H), 1.99 (t, 2H), 3.93 (s, 3H), 4.13 (t, 2H), 4.19 (s, 1H), 7.19 (m,2H), 7.40 (t, 1H), 7.81 (s, 1H), 7.88 (d, 1H), 7.98 (s, 1H), 8.49 (s,1H), 8.68 (s, 1H), 9.47 (s, 1H), 10.39 (s, 1H).

Example 8 Preparation of7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 12) Step 8a. Ethyl6-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0113-12)

The title compound 0113-12 was prepared as a yellow solid (305 mg, 84%)from compound 0112 from step 5b (247 mg, 0.85 mmol) and ethyl7-bromohepanoate (211 mg, 0.89 mmol) using a procedure similar to thatdescribed for compound 0110-1 (Example 1): LCMS: 448 [M+1]⁺; ¹H NMR(CDCl₃): δ 1.15 (t, J=7.5 Hz, 3H), 1.33-1.60 (m, 6H), 1.81 (m, 2H), 2.28(t, J=7.5 Hz, 2H), 3.92 (s, 3H), 4.03 (q, J=7.2 Hz, 2H), 4.12 (t, J=6.6Hz, 2H), 4.18 (s, 1H), 7.19 (m, 2H), 7.39 (t, J=7.8 Hz, 1H), 7.80 (s,1H), 7.89 (d, J=8.1 Hz, 1H), 7.97 (s, 1H), 8.48 (s, 1H), 9.44 (s, 1H).

Step 8b.7-(4-(3-Ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 12)

The title compound 12 was prepared as a grey solid (100 mg, 41%) fromcompound 0113-12 (250 mg, 0.56 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): m.p. 171.8˜177.2° C. (dec); LCMS:435 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.36 (m, 2H), 1.52 (m, 4H), 1.83 (m,2H), 1.97 (m, 2H), 3.94 (s, 3H), 4.14 (t, J=6.3 Hz, 2H), 4.20 (s, 1H),7.21 (m, 2H), 7.41 (t, J=8.1 Hz, 1H), 7.83 (s, 1H), 7.90 (d, J=8.1 Hz,1H), 8.00 (s, 1H), 8.50 (s, 1H), 8.66 (s, 1H), 9.48 (s, 1H), 10.35 (s,1H).

Example 8 (Method 2): Preparation of7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 12) Step 8a′. Ethyl 3-hydroxy-4-methoxybenzoate (Compound0402-12)

To a solution of ethyl 3,4-dihydroxybenzoate 0401 (12.52 g, 68.7 mmol)in DMF (50 mL) was added potassium carbonate (9.48 g, 68.7 mmol). Afterthe mixture was stirred for 15 minutes, a solution of iodomethane (9.755g, 68.7 mmol) in DMF (10 mL) was added dropwise. The reaction mixturewas stirred at 20° C. for 24 hours. After reaction the mixture wasfiltered, and the filtrate was concentrated. The residue was dissolvedin dichloromethane and washed with brine. The organic phase was driedover sodium sulfate, filtered and concentrated in vacuo to give crudeproduct. The crude product was purified by column chromatography to givethe title compound 0402-12 as a white solid (7.1 g, 53%): LCMS: 197[M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.29 (t, J=6.6 Hz, 3H), 3.83 (s, 3H), 4.25(q, J=6.6 Hz, 2H), 7.00 (d, J=8.4 Hz, 1H), 7.38 (d, J=1.8 Hz, 1H), 7.43(dd, J=8.4 Hz, 2.1 Hz, 1H), 9.36 (s, 1H).

Step 8b′. Ethyl 3-(7-ethoxy-7-oxoheptyloxy)-4-methoxybenzoate (Compound0403-12)

A mixture of compound 0402-12 (6.34 g, 32.3 mmol), ethyl7-bromoheptanoate (7.66 g, 32.3 mmol) and potassium carbonate (13.38 g,96.9 mmol) in DMF (80 mL) was stirred at 60° C. for 3 hours. Afterreaction the mixture was filtrated. The filtrate was concentrated invacuo and the residue was dissolved in dichloromethane and washed withbrine twice. The organic phase was dried over sodium sulfate, filteredand concentrated to give the title product 0403-12 as a white solid(9.87 g, 86.7%): LCMS: 353 [M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.17 (t, J=6.9Hz, 3H), 1.31 (t, J=7.2 Hz, 3H) 1.39 (m, 4H), 1.54 (m, 2H), 1.72 (m,2H), 2.29 (t, J=7.2 Hz, 2H), 3.83 (s, 3H), 3.98 (t, J=7.2 Hz, 2H), 4.06(q, J=6.9 Hz, 2H), 4.29 (q, J=7.2 Hz, 2H), 7.06 (d, J=8.4 Hz, 1H), 7.42(d, J=1.8 Hz, 1H), 7.57 (dd, J=8.4 Hz, 1.8 Hz, 1H).

Step 8c′. Ethyl 5-(7-ethoxy-7-oxoheptyloxy)-4-methoxy-2-nitrobenzoate(Compound 0404-12)

Compound 0403-12 (9.87 g, 28.0 mmol) was dissolved in acetic acid (20mL) and stirred at 20° C. Fuming nitric acid (17.66 g, 280.0 mmol) wasadded slowly dropwise. The mixture was stirred at 20° C. for 1 hour.After reaction the mixture was poured into ice-water and extracted withdichloromethane twice. The combined organic phase was washed with brine,aqueous NaHCO₃ solution and brine. The combined organic phase was driedover sodium sulfate, filtered and concentrated to give the title product0404-12 as a yellow solid (10.75 g, 96.4%): LCMS: 398 [M+1]⁺, ¹H NMR(DMSO-d₆): δ 1.17 (t, J=7.2 Hz, 3H), 1.27 (t, J=7.2 Hz, 3H), 1.38 (m,4H), 1.53 (m, 2H), 1.74 (m, 2H), 2.29 (t, J=7.2 Hz, 2H), 3.91 (s, 3H),4.03 (q, J=7.2 Hz, 2H), 4.08 (t, J=6.3 Hz, 2H), 4.30 (q, J=7.2 Hz, 2H),7.29 (s, 1H), 7.63 (s, 1H).

Step 8d′. Ethyl 2-amino-5-(7-ethoxy-7-oxoheptyloxy)-4-methoxybenzoate(Compound 0405-12)

A mixture of 0404-12 (10.75 g 27.0 mmol), ethanol (120 mL), water (40mL) and hydrogen chloride (4 mL) was stirred to form a clear solution.The iron powder (15.16 g, 27.0 mmol) was added batchwise. The mixturewas stirred at reflux for 30 min, and was then cooled to roomtemperature, adjusted pH to 8 with 10% sodium hydroxide solution, andfiltered. The filtrate was concentrated to remove ethanol and extractedwith dichloromethane twice. The combined organic phase was washed withbrine and dried over sodium sulfate, filtered and concentrated to givethe title product 0405-12 as a yellow solid (8.71 g, 87.8%): LCMS: 368[M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.17 (t, J=7.2 Hz, 3H), 1.28 (t, J=7.2 Hz,3H), 1.37 (m, 4H), 1.53 (m, 2H), 1.66 (m, 2H), 2.29 (t, J=7.2 Hz, 2H),3.74 (s, 3H), 3.78 (t, J=6.9 Hz, 2H), 4.06 (q, J=7.2 Hz, 2H), 4.22 (q,J=7.2 Hz, 2H), 6.35 (s, 1H), 6.44 (s, 2H), 7.15 (s, 1H).

Step 8e′. Ethyl7-(7-methoxy-4-oxo-3,4-dihydroquinazolin-6-yloxy)heptanoate (Compound0406-12)

A mixture of compound 0405-12 (8.71 g, 23.7 mmol), ammonium formate(1.48 g, 23.7 mmol) and formamide (40 mL) was stirred at 180° C. for 3hours. After reaction the mixture was cooled to room temperature. Theformamide was removed under reduce pressure, and the residue wasdissolved in dichloromethane and washed with brine. The organic phasewas dried over sodium sulfate, filtered and concentrated to give thetitle product 0406-12 as a pale white solid (8.18 g, 99%): LCMS: 349[M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.17 (t, J=6.9 Hz, 3H), 1.38 (m, 4H), 1.55(m, 2H), 1.75 (m, 2H), 2.29 (t, J=7.2 Hz, 2H), 3.90 (s, 3H), 4.05 (m,4H), 7.13 (s, 1H), 7.42 (s, 1H), 7.97 (d, J=3.6 Hz, 1H), 12.07 (s, 1H).

Step 8f′. Ethyl 7-(4-chloro-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0407-12)

A mixture of product 0406-12 (8.18 g, 23.5 mmol) and phosphoryltrichloride (50 mL) was stirred at reflux for 4 hours. After reactionthe excessive phosphoryl trichloride was removed under reduced pressure.The residue was dissolved in dichloromethane and washed with water,aqueous NaHCO₃ solution and brine. The organic phase was dried oversodium sulfate, filtered and concentrated to give the title product0407-12 as a yellow solid (5.93 g, 69.7%): LCMS: 367 [M+1]⁺, ¹H NMR(DMSO-d₆): δ 1.17 (t, J=6.9 Hz, 3H), 1.38 (m, 4H), 1.54 (m, 2H), 1.81(m, 2H), 2.30 (t, J=7.2 Hz, 2H), 4.02 (s, 3H), 4.06 (q, J=6.9 Hz, 2H),4.18 (t, J=6.3 Hz, 2H), 7.37 (s, 1H), 7.45 (s, 1H), 8.87 (s, 1H).

Step 8g′. Ethyl7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0408-12)

A mixture of product 0407-12 (5.93 g, 16.4 mmol) and3-ethynylbenzenamine (1.92 g, 16.4 mmol) in isopropanol (80 mL) wasstirred at reflux 4 hours. After reaction the mixture was cooled to roomtemperature and resulting precipitate was isolated, washed withisopropanol and ether, and dried to give the title compound 0408-12 as ayellow solid (4.93 g, 67.1%): LCMS: 448 [M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.16(t, J=7.2 Hz, 3H), 1.36-1.59 (m, 6H), 1.80 (m, 2H), 2.29 (t, J=7.2 Hz,2H), 3.93 (s, 3H), 4.04 (q, J=6.9 Hz, 2H), 4.13 (t, J=6.6 Hz, 2H), 4.19(s, 1H), 7.20 (m, 2H), 7.39 (t, J=7.8 Hz, 1H), 7.81 (s, 1H), 7.89 (d,J=8.4 Hz, 1H), 7.97 (s, 1H), 8.48 (s, 1H), 9.45 (s, 1H).

Step 8h′.7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 12)

The freshly prepared hydroxylamine solution (30 mL, 110 mmol) was placedin 50 mL flask. Compound 0408-12 (4.93 g, 11.0 mmol) was added to thissolution and stirred at 25° C. for 24 hours. After reaction the mixturewas neutralized with acetic acid, and the resulting precipitate wasisolated, washed with water, and dried to give the title compound 12 asa white solid (3.99 g, 83.6%): mp 174.1˜177.2° C. LCMS: 435 [M+1]⁺, ¹HNMR (DMSO-d₆): δ 1.36 (m, 2H), 1.52 (m, 4H), 1.83 (m, 2H), 1.98 (m, 2H),3.94 (s, 3H), 4.14 (t, J=6.6 Hz, 2H), 4.20 (s, 1H), 7.21 (m, 2H), 7.41(t, J=7.8 Hz, 1H), 7.80 (s, 1H), 7.90 (d, J=7.8 Hz, 1H), 8.00 (s, 1H),8.50 (s, 1H), 8.66 (s, 1H), 9.48 (s, 1H), 10.35 (s, 1H).

Purification by Crystallization:

To a 30 L 4-necked flask, was added crude product (1.3 kg, KF=11%),followed by THF (26 L). The solution was heated to reflux until a nearlyclear solution. The solution was filtered at 45-55° C., and the filtratewas charged into a 30 L reactor, with stirring speed at 350 (r.m.p),IT=40-42° C. The solution was gradually cooled to 30° C. (during 6-8hours). Solids appeared at 32-33° C., stirred at this temperature for6-8 hours. Then the solution was cooled to 15-25° C. (During 4-5 hours).The solids were collected by filtration, and rinsed with THF (5 L). Thedamp solids were dried under vacuum at 35-40° C. for 15 h, to givecrystalline compound 12 (0.9 Kg, 2.07 mol. Yield 69.2%).

Note (after vacuum drying, there is always about 10% of THF contained inthe compound, pulping in ethyl acetate will remove THF).

Pulping in Ethyl Acetate to Remove THF:

The above solids (0.9 kg) were charged into 30 L 4-necked flack, addedethyl acetate (9 ml), the suspension was stirred at 25-30° C. for 1hour, then was filtered, and rinsed with ethyl acetate (2×2 L). The dampsolids were vacuum dried at 60-70° C. for 20 h to give crystallinecompound 12 (0.86 kg). GC check no THF contained.

Example 9 Preparation of2-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-hydroxyacetamide(Compound 13) Step 9a. 6-Hydroxyquinazolin-4(3H)-one (compound 0202)

To a solution of 2-amino-5-hydroxybenzoic acid 0201 (30.6 g, 0.2 mol) informamide was stirred and heated to 190° C. for 0.5 h. The mixture wasallowed to cool to room temperature. The precipitate was isolated,washed with ether and dried to obtain title compound 0202 (32 g, brownsolid, yield: 99%): LC-MS m/z 163 [M+1]; ¹H NMR (DMSO) δ7.25 (dd, 1H),7.40 (d, 1H), 7.46 (d, 1H), 7.88 (s, 1H).

Step 9b. 3,4-Dihydro-4-oxoquinazolin-6-yl acetate (Compound 0203)

A mixture of compound 0202 (30.0 g, 0.185 mol) and pyridine (35 ml) inacetic anhydride (275 ml) was stirred and heated at 100° C. for 2 hours.The reaction was poured into a mixture of ice and water (500 ml). Theprecipitate was isolated, washed with water and dried to obtain thetitle compound 0203 (24 g, pale white solid, yield: 61%): LC-MS m/z 205[M+1]; 1H-NMR (DMSO) δ 2.32 (s, 3H), 7.50 (dd, 1H), 7.80 (d, 1H), 7.98(s, 1H), 8.02 (s, 1H).

Step 9c. 4-Chloroquinazolin-6-yl acetate (Compound 0204)

A mixture of compound 0203 (20.0 g, 0.1 mol) in POCl₃ (150 ml) wasstirred and heated to reflux for 2 hours. The reaction was evaporatedand the residue was partitioned between ethyl acetate and a saturatedaqueous NaHCO₃ solution. The organic phase was washed with water, driedover Na₂SO₄ and evaporated. The mixture was purified by columnchromatography (silica gel, elution: 1:2=ethyl acetate/petroleum) toobtained the title compound 0204 (7.5 g, white solid, yield: 35%): LC-MSm/z 223 [M+1]; ¹H-NMR (CDCl₃) δ2.40 (s, 3H), 7.74 (dd, 1H), 8.00 (d,1H), 8.09 (d, 1H), 9.05 (s, 1H).

Step 9d. 4-(3-Chloro-4-fluorophenylamino)quinazolin-6-yl acetate(Compound 0207)

A mixture of 0204 (1.0 g, 4.5 mmol) and 3-chloro-4-fluorobenzenamine0205 (0.7 g, 5.0 mmol) in isopropanol (45 ml) was stirred and heated at90° C. for 1 hours. The reaction was cooled to room temperature and theprecipitate was isolated. The solid was washed in turn with isopropanoland methanol, dried to provide the title compound 0207 (1.3 g, paleyellow solid, yield: 87%): LC-MS m/z 332 [M+1]; 1H-NMR (DMSO) δ2.37 (s,3H), 7.54 (t, 1H), 7.75 (m, 1H), 7.94 (dd, 1H), 7.99 (s, 1H), 8.02 (m,1H), 8.64 (s, 1H), 8.95 (s, 1H).

Step 9e. 4-(3-Chloro-4-fluorophenylamino)quinazolin-6-ol (Compound 0209)

A mixture of 0207 (0.8 g, 2.6 mmol) and lithium hydroxide monohydrate(0.13 g, 3.2 mmol) in methanol (10 ml)/water (15 ml) was stirred at roomtemperature for 1 hour. The pH was adjusted to 4 with acetic acid andfiltered. The collected yellow solid was washed by water and dried toobtained title compound 0209 (0.6 g, yellow solid, yield: 88%): LC-MSm/z 290 [M+1]; ¹H-NMR (DMSO) δ7.42 (s, 1H), 7.45 (m, 1H), 7.70 (d, 1H),7.76 (s, 1H), 7.86 (m, 1H), 8.24 (q, 1H), 8.48 (s, 1H), 9.61 (s, 1H).

Step 9f. Ethyl2-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)acetate (Compound0210-13)

A mixture of 0209 (0.2 g, 0.77 mmol), ethyl 3-bromopropanoate (0.14 g,0.85 mmol) and K₂CO₃ (0.8 g, 5.8 mmol) in DMF (15 ml) was stirred andheated to 80° C. for 2 hours. The reaction was filtered and the filtratewas evaporated. The resulting solid was washed with ether to obtain thetitle compound 0210-13 (0.2 g, yellow solid, yield: 75%): mp 161-163°C.; LC-MS m/z 376 [M+1]; 1H-NMR (DMSO) δ1.20 (t, 3H), 4.20 (q, 2H), 4.96(s, 2H), 7.45 (t, 1H), 7.55 (dd, 1H), 7.78 (m, 2H), 7.94 (d, 1H), 8.16(dd, 1H), 8.54 (s, 1H), 9.69 (s, 1H).

Step 9g.2-(4-(3-Chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-hydroxyacetamide(Compound 13)

To a stirred solution of hydroxyamine hydrochloride (4.67 g, 67 mmol) inmethanol (24 ml) at 0° C. was added a solution of potassium hydroxide(5.61 g, 100 mmol) in methanol (14 ml). After addition, the mixture wasstirred for 30 minutes at 0° C., and was allowed to stand at lowtemperature. The resulting precipitate was isolated, and the solutionwas prepared to give free hydroxyamine.

Take above solution (1.4 ml, 2.4 mmol) into 5 ml flask. Compound 0210-13(0.1 g, 0.29 mmol) was added into this solution and stirred at 0° C. for10 minutes, and then allowed to warm to room temperature. The reactionprocess was monitored by TLC. The mixture was adjusted pH to 6 withacetic acid and then concentrated under reduce pressure. The residue waspurified by preparation HPLC eluted by methanol/water. The bandcontaining the product was collected. The solvent was evaporated toobtain title compound 13 (30 mg, yellow solid, yield: 29%): LC-MS m/z363 [M+1]; 1H-NMR (DMSO) δ4.64 (s, 2H), 7.46 (t, 1H), 7.58 (d, 1H), 7.79(d, 2H), 7.7 (s, 1H), 8.11 (s, 1H), 8.52 (s, 1H), 9.02 (s, 1H), 9.67 (s,1H), 10.96 (s, 1H).

Example 10 Preparation of4-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-Hydroxybutanamide(Compound 15)

The title compound 15 was prepared (20 mg) from compound 0209 from step9e and ethyl 4-bromobutanoate using a procedure similar to thatdescribed for compound 13 (Example 9): mp 128-132° C.; LC-MS m/z 391[M+1]; ¹H-NMR (DMSO+D2O) δ2.05 (m, 2H), 2.24 (t, 2H), 4.21 (t, 2H) 7.46(t, 1H), 7.54 (dd, 1H), 7.65 (m, 1H), 7.76 (d, 1H), 7.82 (m 1H), 7.99(m, 1H), 8.43 (s, 1H).

Example 11 Preparation of6-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-hydroxyhexanamide (Compound 17) Step 11a. Ethyl6-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)hexanoate(Compound 0210-17)

The title compound 0210-17 (0.2 g) was prepared from compound 02094-(3-chloro-4-fluorophenylamino)quinazolin-6-ol and ethyl6-bromohexanoate using a procedure similar to that described forcompound 0210-13 (Example 9): LC-MS m/z 433 [M+1], ¹H-NMR (DMSO) δ1.13(t, 3H), 1.45 (m, 2H), 1.60 (m, 2H) 1.76 (m, 2H), 2.30 (t, 2H), 4.05 (q,2H), 4.11 (t, 2H), 7.41 (d, 1H), 7.45 (dd, 1H), 7.68 (d, 1H), 7.80 (m,1H), 7.86 (m, 1H), 8.13 (dd, 1H), 8.48 (s, 1H).

Step 11b.6-(4-(3-Chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-hydroxyhexanamide(Compound 17)

The title compound 17 (30 mg) was prepared from compound 0210-17 using aprocedure similar to that described for compound 13 (Example 9): LC-MS[M+1] 419 ¹H-NMR (DMSO) δ1.28 (m, 2H), 1.60 (m, 2H) 1.73 (m, 2H), 2.05(t, 2H), 4.17 (t, 2H), 7.25 (d, 1H), 7.47 (t, 1H), 7.55 (dd, 1H) 7.76(d, 1H) 7.73 (m, 1H), 8.05 (m, 1H), 8.48 (s, 1H).

Example 12 Preparation of7-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 18) Step 12a. Ethyl7-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)heptanoate(Compound 0210-18)

The title compound 0210-18 (0.2 g) was prepared from compound 2-64-(3-chloro-4-fluorophenylamino)quinazolin-6-ol (0209) of step 9e andethyl 7-bromoheptanoate using a procedure similar to that described forcompound 0210-13 (Example 9): LC-MS m/z 420 [M+1], ¹H-NMR (DMSO) δ1.13(t, 3H), 1.36 (m, 2H), 1.46 (m, 2H), 1.54 (m, 2H), 1.78 (m, 2H), 2.27(t, 2H), 4.05 (q, 2H), 4.11 (t, 2H), 7.41 (d, 1H), 7.47 (dd, 1H), 7.70(d, 1H), 7.81 (m, 1H), 7.84 (m, 1H), 8.13 (dd, 1H), 8.50 (s, 1H).

Step 12b.7-(4-(3-Chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 18)

The title compound 18 (20 mg) was prepared from compound ethyl7-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy-heptanoate(0210-18) using a procedure similar to that described for compound 13(Example 9): LC-MS m/z 433 [M+1], mp 145-149° C., ¹H-NMR (DMSO) δ1.32(m, 2H), 1.47 (m, 4H) 1.88 (m, 2H), 1.94 (t, 2H), 4.12 (t, 2H), 7.43 (t,1H), 7.51 (dd, 1H), 7.71 (d, 1H) 7.80 (m, 1H) 7.86 (d, 1H), 8.15 (dd,1H), 8.51 (s, 1H).

Example 13 Preparation of2-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)-N-hydroxyacetamide(Compound 19) Step 13a. 4-(3-Ethynylphenylamino)quinazolin-6-yl acetate(Compound 0208)

The title compound 0208 (0.8 g, yield: 73%) was prepared from4-chloroquinazolin-6-yl acetate 0204 and 3-ethynylbenzenamine 0206 usinga procedure similar to that described for compound 0207 (Example 9):LC-MS m/z 304 [M+1], ¹H-NMR (DMSO) δ2.36 (s, 3H), 4.26 (s, 1H), 7.43 (d,1H), 7.53 (t, 1H), 7.77 (d, 1H), 7.95 (m, 2H), 8.02 (d, 1H), 8.71 (s,1H), 8.96 (s, 1H).

Step 13b. 4-(3-Ethynylphenylamino)quinazolin-6-ol (Compound 0211)

The title compound 0211 (0.6 g, yield: 88%) was prepared using aprocedure similar to that described for compound 0209 (Example 9): LC-MSm/z 262 [M+1], 1H-NMR (DMSO) δ4.17 (s, 1H), 7.19 (d, 1H), 7.36 (t, 1H),7.43 (dd, 1H, 7.65 (d, 1H), 0.82 (d, 1H), 0.95 (d, 1H), 8.10 (s, 1H),0.48 (s, 1H).

Step 13c. Ethyl 2-(4-(3-Ethynylphenylamino)quinazolin-6-yloxy)acetate(Compound 0212-19)

The title compound 0212-19 (0.2 g, yield: 75%) was prepared from4-(3-ethynylphenylamino)quinazolin-6-ol 0211 and ethyl 2-bromoacetateusing a procedure similar to that described for compound 0210-13(Example 9): LC-MS m/z 322 [M+1], mp 181-182° C.) ¹H-NMR (DMSO) δ1.28(t, 3H), 4.20 (q, 2H), 4.25 (s, 1H), 4.32 (s, 2H), 7.23 (d, 1H), 7.41(t, 1H), 7.57 (dd, 1H), 7.74 (d, 1H), 7.91 (d, 1H), 7.95 (m, 1H), 8.10(s, 1H), 8.48 (s, 1H).

Step 13d.2-(4-(3-Ethynylphenylamino)quinazolin-6-yloxy)-N-hydroxyacetamide(Compound 19)

The title compound 12 (40 mg) was prepared from ethyl2-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)acetate 0212-19 using aprocedure similar to that described for compound 13 (Example 9): LC-MSm/z 335 [M+1], mp: 189-191° C., ¹H-NMR (DMSO) δ4.27 (s, 1H), 4.69 (s,2H), 7.39 (d, 1H), 7.49 (t, 1H), 7.76 (m, 2H), 7.83 (m, 2H), 7.88 (s,1H), 8.10 (s, 1H), 8.82 (m, 1H).

Example 14 Preparation of4-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)-N-hydroxybutanamide(Compound 21) Step 14a. Ethyl4-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)butanoate (Compound0212-21)

The title compound 0212-21 (0.2 g, 78%) was prepared from compound4-(3-ethynylphenylamino)quinazolin-6-ol (0211) and ethyl4-bromobutanoate using a procedure similar to that described forcompound 0210-13 (Example 9): LC-MS m/z 376 [M+1], ¹H-NMR (DMSO) δ1.12(t, 3H), 1.79 (m, 2H), 2.32 (t, 2H), 4.04 (q, 2H), 4.16 (t, 2H), 4.21(s, 1H), 7.02 (dd, 1H), 7.21 (d, 1H), 7.39 (dd, 1H), 7.70 (t, 1H), 7.88(s, 1H), 8.00 (m, 1H), 8.51 (s, 1H), 8.65 (s, 1H).

Step 14b.4-(4-(3-Ethynylphenylamino)quinazolin-6-yloxy)-N-hydroxybutanamide(Compound 21)

The title compound 21 (50 mg) was prepared from ethyl4-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)butanoate (0212-21) usinga procedure similar to that described for compound 13 (Example 9): LC-MSm/z 363 [M+1], mp 182-186° C., ¹H-NMR (DMSO) δ2.02 (m, 2H), 2.20 (t,2H), 4.16 (t, 2H), 4.20 (s, 1H), 7.24 (d, 1H), 7.43 (t, 1H), 7.52 (dd,1H), 7.75 (d, 1H), 7.94 (m, 2H), 8.06 (s, 1H), 8.53 (s, 1H).

Example 15 Preparation of6-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)-N-hydroxyhexanamide(Compound 23) Step 15a.6-(4-(3-Ethynylphenylamino)quinazolin-6-yloxy)hexanoate (Compound0212-23)

The title compound ethyl6-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)hexanoate (0212-23) (0.3g, 64%) was prepared from compound4-(3-ethynylphenylamino)quinazolin-6-ol (0211) and ethyl6-bromohexanoate using a procedure similar to that described forcompound 0210-13 (Example 9): LC-MS m/z 404 [M+1].

Step 15b.6-(4-(3-Ethynylphenylamino)quinazolin-6-yloxy)-N-hydroxyhexanamide(Compound 23)

The title compound 23 (50 mg) was prepared from ethyl6-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)hexanoate (0212-23) usinga procedure similar to that described for compound 13 (Example 9): LC-MSm/z 391 [M+1], mp 176-182° C., ¹H-NMR (DMSO) δ1.46 (m, 2H), 1.60 (m,2H), 1.81 (m, 2H), 2.00 (t, 2H), 4.15 (t, 2H), 4.20 (s, 1H), 7.24 (d,1H), 7.43 (t, 1H), 7.52 (dd, 1H), 7.72 (d, 1H), 7.92 (m, 2H), 8.04 (s,1H), 8.53 (s, 1H).

Example 164-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxybutanamide(Compound 4) Step 16a. Ethyl4-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)butanoate (Compound 0110-4)

The title compound 0110-4 was prepared as a yellow solid (600 mg, 88.4%)from compound 0109 from step 1f (500 mg, 1.56 mmol) and methyl5-bromopentanoate (320 mg, 1.64 mmol) using a procedure similar to thatdescribed for compound 0110-1 (Example 1): LCMS: 434 [M+1]⁺; ¹H NMR(CDCl₃): δ 1.80˜1.97 (m, 4H), 2.48 (t, J=6.6 Hz, 2H), 3.67 (s, 3H), 3.97(s, 3H), 4.18 (t, J=7.2 Hz, 2H), 7.14 (t, J=8.7 Hz, 1H), 7.24 (s, 1H),7.29 (s, 1H), 7.66˜7.11 (m, 1H), 7.96 (dd, J=6.9 Hz, 2.7 Hz, 1H), 8.03(s, 1H), 8.66 (s, 1H).

Step 16b.4-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-Nhydroxybutanamide (Compound 4)

The title compound 4 was prepared as a white solid (140 mg, 35%) formcompound 0110-4 (400 mg, 0.92 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: 435 [M+1]⁺; ¹H NMR(DMSO-d⁶): δ 1.69˜1.84 (m, 4H), 2.07 (t, J=6.6 Hz, 2H), 3.94 (s, 3H),4.15 (t, J=6.0 Hz, 2H), 7.21 (s, 1H), 7.45 (t, J=9.0 Hz, 1H), 7.78˜7.83(m, 2H), 8.13 (dd, J=6.9 Hz, 2.4 Hz, 1H), 8.03 (s, 1H), 8.50 (s, 1H),8.72 (s, 1H), 9.54 (s, 1H), 10.41 (s, 1H).

Example 175-(4-(3-Ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxypentanamide(Compound 10) Step 17a. Methyl5-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)pentanoate(Compound 0113-10)

The title compound 0113-10 was prepared as a yellow solid (500 mg, 72%)from compound 0112 (500 mg, 1.7 mmol) and methyl 5-bromopentanoate (211mg, 0.89 mmol) using a procedure similar to that described for compound0110-1 (Example 1): LCMS: 406 [M+1]⁺.

Step 17b.5-(4-(3-Ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxypentanamide(Compound 10)

The title compound 10 was prepared as a white solid (200 mg, 40%) fromcompound 0113-10 (500 mg, 1.23 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: 407 [M+1]⁺; ¹H NMR(DMSO-d₆): δ 1.71˜1.85 (m, 4H), 2.07 (t, J=7.2 Hz, 2H), 3.93 (s, 3H),4.16 (t, J=6.3 Hz, 2H), 4.20 (s, 1H), 7.19 (m, 2H), 7.41 (t, J=8.1 Hz,1H), 7.84 (s, 1H), 7.90 (dd, J=8.4 Hz, 1.2 Hz, 1H), 8.00 (t, J=1.8 Hz,1H), 8.50 (s, 1H), 8.72 (s, 1H), 9.48 (s, 1H), 10.40 (s, 1H).

Example 18 Preparation of5-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-hydroxypentanamide(Compound 16) Step 18a. ethyl5-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)pentanoate(Compound 0210-16)

The title compound 0210-16 (0.2 g, 68%) was prepared from compound 02094-(3-chloro-4-fluorophenylamino)quinazolin-6-ol (0.2 g, 0.69 mmol) andmethyl 5-bromopentanoate (0.14 g, 0.69 mmol) using a procedure similarto that described for compound 0210-13 (Example 9): LCMS 376 [M+1]⁺.

Step 18b.5-(4-(3-chloro-4-fluorophenylamino)quinazolin-6-yloxy)-N-hydroxypentanamide(Compound 16)

The title compound 16 (24 mg, 67%) was prepared from compound 0210-16(37 mg, 0.09 mmol) using a procedure similar to that described forcompound 13 (Example 9): mp: 85.9° C.; LCMS 405 [M+1]⁺, ¹H NMR (DMSO-d₆)δ 1.74 (m, 4H), 2.04 (t, J=7.5 Hz, 2H), 4.14 (t, J=6 Hz, 2H), 7.44 (t,J=9 Hz, 1H), 7.51 (dd, J=9 Hz, J=2.4 Hz, 1H), 7.73 (d, J=8.7 Hz, 1H),7.82 (m, 1H), 7.88 (d, J=2.4, 1H) 8.16 (dd, J=6.9 Hz, J=2.7 Hz 1H), 8.52(s, 1H), 8.69 (s, 1H), 9.67 (s, 1H), 10.38 (s, 1H).

Example 19 Preparation of7-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 24) Step 19a. Ethyl7-(4-(3-ethynylphenylamino)quinazolin-6-yloxy)heptanoate (Compound0212-24)

The title compound 0212-24 (0.21 g, 58%) was prepared from compound4-(3-ethynylphenylamino)quinazolin-6-ol (0211) (0.23 g, 0.86 mmol) andethyl 7-bromoheptanoate (0.20 g, 0.86 mmol) using a procedure similar tothat described for compound 0210-13 (Example 9): LCMS 418 [M+1]⁺.

Step 19b.7-(4-(3-Ethynylphenylamino)quinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 24)

The title compound 24 (50 mg, 42%) was prepared from compound 0212-24(123 mg, 0.29 mmol) using a procedure similar to that described forcompound 13 (Example 9): LCMS 405 [M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.44 (m,2H), 1.48 (m, 2H), 1.59 (m, 2H), 1.67 (m, 2H), 2.11 (t, J=7.2 Hz, 2H),3.50 (s, 1H), 4.17 (t, J=6.3 Hz, 2H), 7.28 (d, J=7.5 Hz, 1H), 7.37 (t,J=6.9 Hz, 1H), 7.48 (d, J=9.0 Hz, 1H), 7.78 (dd, J=21.3 Hz, J=7.8 Hz,1H), 7.93 (s, 1H), 7.92 (m, 2H), 8.45 (s, 1H).

Example 20 Example 1 Synthesis of7-(4-(3-Chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 30) Step 20a. Ethyl 3-hydroxy-4-(2-methoxyethoxy)benzoate(Compound 0402-30)

To a solution of 0401 (1.82 g, 10.0 mmol) in N,N-dimethylformamide (20mL) was added potassium carbonate (1.38 g, 10.0 mmol). The mixture wasstirred for 15 minutes and then a solution of 2-methoxyethyl4-methylbenzenesulfonate (2.30 g, 10.0 mmol) in N,N-dimethylformamide (5mL) was added slowly dropwise. The mixture was stirred 48 hours at roomtemperature and filtered. The filtrate was concentrated in vacuo and theresidue was dissolved in ethyl acetate (30 mL) then the organic layerwas washed with brine (20 mL×3) and dried over sodium sulfate, filteredand evaporated to give the title product 0402-30 as a white solid (1.2g, 50%): LCMS: 241 [M+1]⁺. ¹H NMR (DMSO-d₆) δ 1.26 (t, J=7.5 Hz, 3H),3.65 (m, J=1.5 Hz, 2H), 4.11 (m, J=4.5 Hz, 2H), 4.21 (m, J=4.5 Hz, 2H),7.00 (d, J=9 Hz, 1H), 7.37 (m, J=2 Hz, 2H), 9.40 (s, 1H).

Step 20b. Ethyl 3-(7-ethoxy-7-oxoheptyloxy)-4-(2-methoxyethoxy)benzoate(Compound 0403-30)

Compound 0402-30 (204.0 mg, 0.85 mmol) and ethyl 7-bromoheptanoate(201.0 mg, 0.85 mmol) and potassium carbonate (353.0 mg, 2.50 mmol) inN,N-dimethylformamide (5 mL) was stirred at 60° C. for 3 hours. Themixture was filtrated. The filtrate was concentrated in vacuo and theresidue was dissolved in ethyl acetate (30 mL) then the organic layerwas washed with brine (20 mL×3) and dried over sodium sulfate, filteredand evaporated to give the title product 0403-30 as a yellow solid (325mg, 96%): LCMS: 397 [M+1]⁺.

Step 20c. Ethyl5-(7-ethoxy-7-oxoheptyloxy)-4-(2-methoxyethoxy)-2-nitrobenzoate(Compound 0404-30)

Compound 0403-30 (325.0 mg, 0.82 mmol) was dissolved in acetic acid (2mL) and stirred at room temperature. Then fuming nitric acid (0.39 g,6.0 mmol) was added slowly dropwise. The mixture was stirred at roomtemperature for 2 hours. Poured into ice-water (50 mL) and extractedwith ethyl acetate (20 mL×2). The combined organic layer was washed withaqueous NaHCO₃ solution (10 mL×3) and brine (10 mL×3) and dried oversodium sulfate, filtered and evaporated to give the title product0404-30 as a yellow oil (330 mg, 100%): LCMS: 442 [M+1]⁺.

Step 20d. Ethyl2-amino-5-(7-ethoxy-7-oxoheptyloxy)-4-(2-methoxyethoxy)benzoate(Compound 0405-30)

A mixture of 0404-30 (370.0 mg 0.82 mmol), ethanol (4.4 mL), water (3mL) and hydrogen chloride (0.08 mL) was stirred to form a clearsolution. The powder iron (459.0 mg, 8.2 mmol) was added. The mixturewas stirred at reflux for 30 minutes and cooled to room temperature,adjust pH to 8 with 10% sodium hydroxide solution in ice-water bath. Themixture was filtered and the filtrate was concentrated to remove ethanoland was then extracted whit ethyl acetate (20 mL×2). The combinedorganic layer was washed with brine (10 mL×3) and dried over sodiumsulfate, filtered and evaporated to give the title product 0405-30 as ayellow oil (315 mg, 93%): LCMS: 412 [M+1]⁺.

Step 20e. Ethyl7-(7-(2-methoxyethoxy)-4-oxo-3,4-dihydroquinazolin-6-yloxy)heptanoate(Compound 0406-30)

A mixture of compound 0405-30 (315.0 mg, 0.76 mmol), ammonium formate(48.0 mg, 0.76 mmol) and formamide (2.46 mL) was stirred at 190° C. for3 hours. The reaction mixture was cooled to room temperature. Theformamide was removed under reduce pressure, and the residue wasdissolved in ethyl acetate (30 mL). The organic layer was washed withbrine (10 mL×5) and dried over sodium sulfate, filtered and evaporatedto give the title product 0406-30 as a white solid (235 mg, 98%): LCMS:393 [M+1]⁺.

Step 20f. Ethyl7-(4-chloro-7-(2-methoxyethoxy)quinazolin-6-yloxy)heptanoate (Compound0407-30)

A mixture of product 0406-30 (235.0 mg, 0.6 mmol) and phosphoryltrichloride (3 mL) was stirred at reflux for 4 hours. When a clearsolution was obtained, the excessive phosphoryl trichloride was removedunder reduced pressure. The residue was dissolved in ethyl acetate (30mL) and the organic layer was washed in turn with water (10 mL×2),aqueous NaHCO₃ solution (10 mL×2) and brine (20 mL×1), dried over sodiumsulfate, filtered and evaporated to give the title product 0407-30 as ayellow solid (233 mg, 94%): LCMS: 411 [M+1]⁺.

Step 20g. Ethyl7-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)-quinazolin-6-yloxy)heptanoate(Compound 0408-30)

A mixture of product 0407-30 (117.0 m g, 0.28 mmol) and3-chloro-4-fluorobenzenamine (50.0 mg, 0.34 mmol) in isopropanol (3 mL)was stirred at reflux overnight. The mixture was cooled to roomtemperature and resulting precipitate was isolated, washed withisopropanol and ether. The solid was then dried to give the titlecompound 0408-30 as a yellow solid (102 mg, 70%): LCMS: 520 [M+1]⁺.

Step 20h.7-(4-(3-Chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 30)

The freshly prepared hydroxylamine solution (3 mL, 2.0 mmol) was placedin 25 mL flask. Compound 408-30 (102.0 mg, 0.2 mmol) was added andstirred at room temperature for 24 hours. The mixture was neutralizedwith acetic acid/methanol. The mixture was concentrated under reducepressure. The residue was purified by preparation HPLC to give the titlecompound 30 as a yellow solid (85 mg, 84%): LCMS: 507 [M+1]⁺; ¹H NMR(DMSO-d₆) δ 1.33 (m, 2H), 1.50 (m, 4H), 1.79 (s, 2H), 1.94 (t, 2H), 3.29(s, 3H), 3.72 (s, 2H), 4.11 (s, 2H), 4.25 (s, 2H), 7.19 (s, 1H), 7.42(t, 1H), 7.79 (s, 1H), 8.10 (d, 1H), 8.47 (s, 1H), 8.65 (s, 1H), 9.52(s, 1H), 10.33 (s, 1H).

Example 21 Preparation of7-(4-(3-Ethynylphenylamino)-7-(2-methoxyethoxy)quinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 36) Step 21a. Ethyl7-(4-(3-ethynylphenylamino)-7-(2-methoxyethoxy)quinazolin-6-yloxy)heptanoate(Compound 0408-36)

A mixture of product 0407-30 (102.0 mg, 0.25 mmol) and3-ethynylbenzenamine (35.0 mg, 0.3 mmol) in isopropanol (3 mL) wasstirred at reflux overnight. The mixture was cooled to room temperatureand resulting precipitate was isolated, washed with isopropanol andether. The solid was then dried to give the title compound 0408-36 as ayellow solid (88 mg, 72%): LCMS: 491 [M+1]⁺.

Step 21b.7-(4-(3-Ethynylphenylamino)-7-(2-methoxyethoxy)quinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 36)

The freshly prepared hydroxylamine solution (3 mL, 2 mmol) was placed in25 mL flask. Compound 0408-36 (88.0 mg, 0.18 mmol) was added to thissolution and stirred at room temperature for 24 hours. The mixture wasneutralized with acetic acid/methanol and was concentrated under reducepressure. The residue was purified by preparative HPLC to give the titlecompound 36 as a white solid (40 mg, 47%): LCMS: 479 [M+1]⁻; ¹H NMR(DMSO-d₆) δ 1.33 (m, 2H), 1.50 (m, 4H), 1.79 (s, 1H), 1.94 (t, 2H), 3.72(s, 2H), 4.11 (s, 2H), 4.25 (s, 2H), 7.19 (s, 1H), 7.42 (t, 1H), 7.79(s, 1H), 8.10 (d, 1H), 8.47 (s, 1H), 8.65 (s, 1H), 9.52 (s, 1H), 10.33(s, 1H).

Example 22 Preparation ofN¹-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yl)-N⁵-hydroxyglutaramide(Compound 38) Step 22a. 7-Chloroquinazolin-4(3H)-one (Compound 0302)

A mixture of compound 0301 (17.2 g, 100 mmol) and formamide (20 mL) wasstirred at 130° C. for 30 minutes and to 190° C. for 4 hours. Themixture was allowed to cool to room temperature. It was then poured intoa mixture of ice and water. The precipitate was isolated, washed withwater and dried to give the title compound 0302 (15.8 g, 87.7%). ¹H NMR(DMSO-d₆): δ 7.65 (dd, 1H), 7.72 (d, 1H), 8.12 (d, 1H), 8.36 (s, 1H).

Step 22b. 7-Chloro-6-nitroquinazolin-4(3H)-one (compound 0303)

Compound 0302 (18.0 g, 100 mmol) was added portionwise to a stirredmixture of concentrated sulfuric acid (60 mL) and fuming nitric acid (60mL) which had been cooled to 0° C., the mixture was stirred at ambienttemperature for 1 hour and then heated to 45° C. overnight. The mixturewas poured into the mixture of ice and water. The precipitate wasisolated, washed with water and dried. Recrystallization from aceticacid to give the title compound 0303 (14.1 g, 62.7%). ¹H NMR (DMSO-d₆):δ 8.00 (s, 1H), 8.27 (s, 1H), 8.65 (s, 1H), 12.70 (s, 1H).

Step 22c. 7-Methoxy-6-nitroquinazolin-4(3H)-one (compound 0304)

A mixture of compound 0303 (4.0 g, 18.0 mmol) and sodium (2.4 g, 45mmol) in methanol (50 mL) was heated at 100° C. in a sealed pressurevessel for 20 hours. The solution was neutralized with acetic acid anddiluted with water to give the title compound 0304 (3.0 g, 77%). ¹H NMR(DMSO-d₆): δ 4.10 (s, 3H), 7.40 (s, 1H), 8.24 (s, 1H), 8.50 (s, 1H),12.67 (s, 1H).

Step 22d. 4-Chloro-7-methoxy-6-nitroquinazoline (compound 0305)

Compound 0304 (3.8 g, 17.2 mmol) was suspended in POCl₃ (75 mL), themixture was heated to reflux for 4 hours. The additional POCl₃ wasremoved in a vacuum. The residue was dissolved in a mixture ofdichloromethane (50 mL) and aqueous NaHCO₃ (50 mL). The organic layerwas dried and the solvent was removed to give the title compound 0305(3.4 g, 83%). ¹H NMR (DMSO-d₆): δ 4.05 (s, 3H), 7.44 (s, 1H), 8.27 (s,1H), 8.53 (s, 1H).

Step 22e.N-(3-chloro-4-fluorophenyl)-7-methoxy-6-nitroquinazolin-4-amineHydrochloride (compound 0307)

A mixture of compound 0305 (3.4 g, 14.2 mmol) and3-chloro-4-fluoroaniline (0406) (2.2 g, 15.2 mmol) and isopropanol (120mL) was stirred at reflux for 3 hours. The mixture was cooled to ambienttemperature and the precipitate was isolated, washed with methanol andether and then dried to give the title compound 0307 (4.66 g, 85%). ¹HNMR (DMSO-d₆): δ 4.10 (s, 3H), 7.55 (dd, 2H), 7.74 (m, 1H), 8.07 (dd,1H), 8.90 (s, 1H), 9.55 (s, 1H), 11.6 (s, 1H).

Step 22f.N-(3-chloro-4-fluorophenyl)-7-methoxy-6-nitroquinazolin-4-amine(Compound 0308)

A mixture of compound 0307 (3.5 g, 10.0 mmol) and iron dust (11.2 g,200.0 mmol) and ethanol (100 mL) and concentrated hydrochloric acid (2mL), and water (30 mL) was heated to reflux for 1 hour. Removed irondust by filtration. The filtrate was concentrated to ⅕ volume. Theprecipitate was isolated and dried to give the title compound 0308 (2.2g, 69%). ¹H NMR (DMSO-d₆): δ 3.97 (s, 3H), 5.38 (s, 2H), 7.10 (s, 1H),7.36 (s, 1H), 7.39 (t, 1H), 7.80 (m, 1H), 8.08 (dd, 1H), 8.38 (s, 1H),9.39 (s, 1H).

Step 22g. Methyl3-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-ylcarbamoyl)propanoate(Compound 0310-38)

The compound 0308 (500.0 mg, 1.57 mmol) and triethylamine (165.0 mg,1.65 mmol) was dissolved in dichloromethane (50 mL). The mixture wascooled to 0° C. and the solution of methyl 5-chloro-5-oxopentanoate (270mg, 1.65 mmol) in dichloromethane (5 mL) was added into above mixturedropwise under 0° C. in 20 minutes. The reaction mixture was allowed tostir at ambient temperature for 1 hour. The mixture was washed withwater (50 mL×2) and brine (50 mL). The organic layer was dried overMgSO₄, filtered and concentrated to give the title compound 0310-38 (550mg, 78%), LCMS: 448 [M+1]⁺.

Step 22h.N¹-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yl)-N⁵-hydroxyglutaramide(Compound 38)

To a stirred solution of hydroxylamine hydrochloride (4.67 g, 67 mmol)in methanol (24 mL) at 0° C. was added a solution of potassium hydroxide(5.61 g, 100 mmol) in methanol (14 mL). After addition, the mixture wasstirred for 30 minutes at 0° C. and was allowed to stand at lowtemperature. The resulting precipitate was isolated, and the solutionwas prepared to give free hydroxylamine.

The above freshly prepared hydroxylamine solution (5.6 mL, 10.0 mmol)was placed in 10 mL flask. Compound 0310-38 (550.0 mg, 1.23 mmol) wasadded to this solution and stirred at 0° C. for 10 minutes and wasallowed to warm to room temperature. The reaction process was monitoredby TLC. The mixture was neutralized with acetic acid. The mixture wasconcentrated under reduce pressure. The residue was purified bypreparative HPLC to give the title compound 38 as a grey solid (250 mg,45%): LCMS: 448 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.85 (m, 2H), 2.06 (t, J=7.5Hz, 2H), 2.48 (t, J=7.2 Hz, 2H), 4.00 (s, 3H), 7.24 (s, 1H), 7.42 (t,J=9.0 Hz, 1H), 7.80 (m, 1H), 8.10 (dd, J=7.2 Hz, 2.7 Hz, 1H), 8.52 (s,1H), 8.70 (s, 1H), 8.82 (s, 1H), 9.48 (s, 1H). 9.79 (s, 1H), 10.40 (s,1H).

Example 23 Preparation ofN¹-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yl)-N⁸-hydroxyoctanediamide(Compound 40) Step 23a. Methyl8-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-ylamino)-8-oxooctanoate(Compound 0310-40)

The title compound 0310-40 was prepared as a yellow solid (350 mg, 78%)from compound 0308 (319 mg, 1.0 mmol) and methyl 8-chloro-8-oxooctanoate(227 mg, 1.1 mmol) using a procedure similar to that described forcompound 0310-38 (Example 22): LCMS: 489 [M+1]⁺.

Step 23b.N¹-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yl)-N⁸-hydroxyoctanediamide(Compound 40)

The title compound 40 was prepared as a yellow solid (120 mg, 30%) fromcompound 0310-38 (400 mg, 0.8 mmol) using a procedure similar to thatdescribed for compound 38 (Example 22): LCMS: 490 [M+1]⁺; ¹H NMR(DMSO-d₆): δ 1.29 (m, 4H), 1.48 (m, 2H), 1.59 (m, 2H), 1.93 (t, J=7.2Hz, 2H), 2.45 (t, J=7.2 Hz, 2H), 4.00 (s, 3H), 4.18 (s, 1H), 7.26 (s,1H), 7.41 (t, J=9.0 Hz, 1H), 7.74 (m, 1H), 8.08 (d, J=1.2 Hz, 1H), 8.54(s, 1H), 8.66 (s, 1H), 8.83 (s, 1H), 9.46 (s, 1H), 9.95 (s, 1H), 10.33(s, 1H).

Example 24 Preparation ofN¹-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yl)-N⁵-hydroxyglutaramide(Compound 42) Step 24a.N-(3-ethynylphenyl)-7-methoxy-6-nitroquinazolin-4-amine Hydrochloride(Compound 0307-42)

The title compound 0307-42 was prepared as a yellow solid (4.7 g, 84.5%)from compound 0305 (350 mg, 0.78 mmol) and 3-ethynylbenzenamine (2.34 g,20.0 mmol) using a procedure similar to that described for compound0306-38 (Example 22): LCMS: 321 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 4.11 (s,3H), 4.24 (s, 1H), 7.42 (d, 1H), 7.50 (t, 1H), 7.61 (s, 1H), 7.79 (d,1H), 7.93 (m, 1H), 8.93 (s, 1H), 9.57 (s, 1H), 11.56 (bs, 1H).

Step 24b. N⁴-(3-ethynylphenyl)-7-methoxyquinazoline-4,6-diamine(Compound 0309-42)

The title compound 0309-42 was prepared as a yellow solid (2.0 g, 69%)from compound 0307-42 (3.2 g, 10.0 mmol) using a procedure similar tothat described for compound 0308-38 (Example 22): LCMS: 291 [M+1]⁺; ¹HNMR (DMSO-d₆): δ 3.95 (s, 3H), 4.14 (s, 1H), 5.33 (s, 2H), 7.08 (m, 2H),7.34 (m, 2H), 7.88 (m, 1H), 8.04 (s, 1H), 8.36 (s, 1H), 9.29 (s, 1H).

Step 24c. Methyl5-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-ylamino)-5-oxopentanoate(Compound 0311-42)

The title compound 0311-42 was prepared as a yellow solid (450 mg, 77%)from compound 0309-42 (407 mg, 1.4 mmol) and methyl5-chloro-5-oxopentanoate (254 mg, 1.54 mmol) using a procedure similarto that described for compound 0310-38 (Example 22): LCMS: 419 [M+1]⁺.

Step 24d.N¹-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yl)-N⁵-hydroxyglutaramide(Compound 42)

The title compound 42 was prepared as a yellow solid (100 mg, 47%) fromcompound 0311-42 (211 mg, 0.5 mmol) using a procedure similar to thatdescribed for compound 38 (Example 22).

Example 25 Preparation ofN¹-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yl)-N⁶-hydroxyadipamide(Compound 43) Step 25a. Methyl6-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-ylamino)-6-oxohexanoate(Compound 0311-43)

The title compound 0311-43 was prepared as a yellow solid (530 mg, 71%)from compound 0309-42 (500 mg, 1.72 mmol) and methyl6-chloro-6-oxohexanoate (323 mg, 1.81 mmol) using a procedure similar tothat described for compound 0311-42 (Example 24): LCMS: 433 [M+1]⁺.

Step 25b.N¹-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yl)-N⁶-hydroxyadipamide(Compound 43)

The title compound 43 was prepared as a yellow solid (105 mg, 24%) fromcompound 0311-43 (432 mg, 1.0 mmol) using a procedure similar to thatdescribed for compound 42 (Example 24): m.p.: 191.2˜196.7° C.; LCMS: 434[M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.58 (m, 4H), 1.98 (t, J=6.3 Hz, 2H), 2.44(m, 2H), 3.99 (s, 3H), 4.16 (s, 1H), 7.18 (d, J=7.8 Hz, 1H), 7.25 (s,1H), 7.37 (t, J=8.1 Hz, 1H), 7.84 (d, J=8.4 Hz, 1H), 7.98 (s, 1H), 8.51(s, 1H), 8.66 (s, 1H), 8.82 (s, 1H), 9.42 (s, 1H), 9.73 (s, 1H), 10.35(s, 1H).

Step 26a. Methyl8-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-ylamino)-8-oxooctanoate(Compound 0311-44)

The title compound 0311-44 was prepared as a yellow solid (150 mg, 78%)from compound 0309-42 (120 mg, 0.4 mmol) and methyl8-chloro-8-oxooctanoate (91 mg, 0.44 mmol) using a procedure similar tothat described for compound 0311-42 (Example 24): LCMS: 461 [M+1]⁺.

Step 26b.N¹-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yl)-N⁸-hydroxyoctanediamide(Compound 44)

The title compound 44 was prepared as a yellow solid (30 mg, 20%) fromcompound 0311-44 (150 mg, 0.3 mmol) using a procedure similar to thatdescribed for compound 42 (Example 24): LCMS: 462 [M+1]⁺; ¹H NMR(DMSO-d₆): δ 1.30 (m, 4H), 1.51 (m, 2H), 1.62 (m, 2H), 1.95 (t, J=7.2Hz, 2H), 2.45 (t, J=7.2 Hz, 2H), 4.00 (s, 3H), 4.18 (s, 1H), 7.19 (d,J=7.2 Hz, 1H), 7.26 (s, 1H), 7.38 (t, J=7.8 Hz, 1H), 7.86 (d, J=7.8 Hz,1H), 7.99 (s, 1H), 8.52 (s, 1H), 8.83 (s, 1H), 9.44 (s, 1H).

Example 27 Preparation of(E)-3-(4-(2-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)ethoxy)phenyl)-N-hydroxyacrylamide(Compound 66) Step 27a. (E)-Methyl 3-(4-hydroxyphenyl)acrylate (Compound0501-66)

A mixture of 4-hydroxycinnamic acid (8.2 g, 50 mmol) and a drop of H₂SO₄in methanol (30 mL) was heated to reflux overnight. Then the solvent wasevaporated, the residue was dissolved in ethyl acetate, washed withsaturated NaHCO₃ solution twice, brine, dried over MgSO₄, concentratedto give the title compound 0501-66 as white solid (8.7 g, 98%): LCMS:179 [M+1]⁺.

Step 27b. (E)-Methyl 3-(4-(2-(tosyloxy)ethoxy)phenyl)acrylate (Compound0502-66)

A mixture of compound 0501-66 (5.0 g, 28.0 mmol) and 2-bromoethanol (3.9g, 62.0 mmol) and potassium carbonate in N,N-dimethylformamide wasstirred at 80° C. for 24 hours. The reaction process was monitored byTLC. The mixture was filtrated. The filtrate was concentrated underreduce pressure. The residue was wash with diethyl ether and dried togive (E)-methyl 3-(4-(2-hydroxyethoxy)phenyl)-acrylate as yellow solid(1.6 g, 26.0%): LCMS: 223 [M+1]⁺.

To a mixture of triethylamine (0.3 g, 3 mol) and dichloromethane (20 mL)was added tosyl chloride (285 mg, 1.5 mmol) batchwise and stirred for0.5 hour. Compound (E)-methyl 3-(4-(2-hydroxyethoxy)phenyl)acrylate (333mg, 1.5 mmol) was added into above mixture and heated to reflux for 24hours. The reaction mixture was added saturated ammonium chloridesolution and the organic layer was separated and washed by brine, dried(MgSO₄), evaporated to give compound 0502-66 as white solid (200 mg,36%): LCMS: 377 [M+1]⁺.

Step 27c. (E)-Methyl3-(4-(2-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)ethoxy)phenyl)acrylate(Compound 0503-66)

A mixture of compound 0109 (176 mg, 0.55 mmol) and 0502-66 (152 mg, 0.94mmol) and potassium carbonate in N,N-dimethylformamide was stirred at80° C. for 24 hours. The reaction process was monitored by TLC. Themixture was filtrated. The filtrate was concentrated under reducepressure. The residue was wash with diethyl ether and dried to give thetitle compound 0503-66 as yellow solid (281 mg, 98%): LCMS: 524 [M+1]⁺.

Step 27d.(E)-3-(4-(2-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)ethoxy)phenyl)-N-hydroxyacrylamide(Compound 66)

The title compound 66 was prepared as a white solid (65 mg, 19%) fromcompound 0503-66 (346.0 mg, 0.66 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: 525 [M+1]⁺; ¹H NMR(DMSO-d₆): δ 3.93 (s, 3H), 4.48 (s, 4H), 6.31 (d, J=16.2 Hz, 1H), 7.05(d, J=8.1 Hz, 2H), 7.21 (s, 1H), 7.44 (t, J=9.0 Hz, 1H), 7.52 (d, J=8.1Hz, 2H), 7.78 (d, J=10.2 Hz, 1H), 7.88 (m, 1H), 8.12 (dd, J=6.6 Hz, 2.7Hz, 1H), 8.50 (s, 1H), 8.96 (s, 1H), 8.50 (s, 1H), 9.56 (s, 1H), 10.65(s, 1H).

Example 28 Preparation ofN¹-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-yl)-N⁵-hydroxyglutaramide(Compound 68) Step 28a. 7-(2-Methoxyethoxy)-6-nitroquinazolin-4(3H)-one(compound 0304-68)

Sodium (2.07 g, 90 mmol) was added to 2-methoxyethanol (125 mL) at 0° C.until sodium was dissolved. Compound 0303 (6.77 g, 30.0 mmol) was addedto the solution. The mixture was stirred at 90° C. for 24 hours and wasthen adjusted to pH7 by acetic acid. Water (50 mL) was added to themixture and resulting yellow precipitate was isolated, washed with waterand dried to provide the title compound 0304-68 as a yellow solid (7.003g, 88%): LCMS: 266 [M+1]⁺.

Step 28b. 4-Chloro-7-(2-methoxyethoxy)-6-nitroquinazoline (compound0305-68)

A mixture of product 0304-68 (5.30 g, 20.0 mmol) and phosphoryltrichloride (50 mL) was stirred at reflux for 5 hours. When a clearsolution was obtained, the excessive phosphoryl trichloride was removedunder reduced pressure. The residue was dissolved in ethyl acetate (100mL) and the organic layer was washed in turn with water (30 mL×2),aqueous NaHCO₃ solution (20 mL×2) and brine (20 mL×1), dried over sodiumsulfate, filtered and evaporated to give the title product 0305-68 as ayellow solid (5.31 g, 94%): LCMS: 284 [M+1]⁺.

Step 28c.N-(3-chloro-4-fluorophenyl)-7-(2-methoxyethoxy)-6-nitroquinazolin-4-amine(compound 0306-68)

A mixture of product 0305-68 (5.31 g, 18.7 mmol) and3-chloro-4-fluorobenzenamine (5.45 g, 37.4 mmol) in isopropanol (150 mL)was stirred at reflux overnight. The mixture was cooled to roomtemperature and resulting precipitate was isolated, washed with methanoland ether. The solid was then dried to give the title compound 0306-68as a yellow solid (5.70 g, 77%): LCMS: 393 [M+1]⁺.

Step 28d.N⁴-(3-chloro-4-fluorophenyl)-7-(2-methoxyethoxy)quinazoline-4,6-diamine(compound 0308-68)

A mixture of 0306-68 (5.70 g, 14.5 mmol), ethanol (165 mL), water (43.5mL) and hydrogen chloride (2.9 mL) was stirred to form a clear solution.The powder iron (16.24 g, 290.0 mmol) was added. The mixture was stirredat reflux for 2 hours. Cooled to room temperature, adjusted pH to 11with 10% sodium hydroxide solution in ice-water bath and was filtered.The filtrate was concentrated to remove ethanol and extracted whit ethylacetate (100 mL×2), The combined organic layer was washed with brine (30mL×3) and dried over sodium sulfate, filtered and evaporated to give thetitle product 0308-68 as a yellow solid (4.92 g, 93%): LCMS: 363 [M+1]⁺.

Step 28e. Methyl5-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-ylamino)-5-oxopentanoate(Compound 0310-68)

The methyl 5-chloro-5-oxopentanoate (0.198 g, 1.2 mmol) was added to asolution of compound 0308-68 (0.22 g, 0.6 mmol) in 30 mL ofdichloromethane and triethylamine (0.48 g, 4.8 mmol). The mixture wasstirred for 2 hours at 0° C. The reaction mixture was then washed withwater and dried over sodium sulfate, filtered and evaporated to give thetitle product 0310-68 as a brown oil (270 mg, 92%): LCMS: 491 [M+1]⁺.

Step 28f.N¹-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-yl)-N⁵-hydroxyglutaramide(Compound 68)

To a stirred solution of hydroxylamine hydrochloride (4.67 g, 67 mmol)in methanol (24 mL) at 0° C. was added a solution of potassium hydroxide(5.61 g, 100 mmol) in methanol (14 mL). After addition, the mixture wasstirred for 30 minutes at 0° C. and was allowed to stand at lowtemperature. The resulting precipitate was isolated, and the solutionwas prepared to give free hydroxylamine.

The above freshly prepared hydroxylamine solution (6 mL, 4.0 mmol) wasplaced in 25 mL flask. Compound 0310-68 (270 mg, 0.55 mmol) was added tothis solution and stirred at room temperature for 4 hours. The mixturewas neutralized with acetic acid/methanol. The mixture was concentratedunder reduce pressure. The residue was purified by preparative HPLC togive the title compound 68 as a yellow solid (220 mg, 75%): LCMS: 492[M+1]⁺; ¹H NMR (DMSO-d₆): 1.83 (m, J=7.5 Hz, 2H), 2.05 (t, J=7.2 Hz,2H), 2.43 (t, J=6.9 Hz, 2H), 3.31 (s, 3H), 3.76 (t, J=4.5 Hz, 2H), 4.32(t, J=4.2 Hz, 2H), 7.28 (s, 1H), 7.40 (t, J=9 Hz, 1H), 7.77 (m, 1H),8.10 (m, J=2.1 Hz, 1H), 8.50 (s, 1H), 8.67 (s, 1H), 8.752 (s, 1H), 9.33(s, 1H), 9.77 (s, 1H), 10.38 (s, 1H).

Example 29 Preparation ofN¹-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-yl)-N⁶-hydroxyadipamide(Compound 69) Step 29a. Methyl6-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-ylamino)-6-oxohexanoate(Compound 0310-69)

The methyl 6-chloro-6-oxohexanoate (0.36 g, 1.76 mmol) was added to asolution of compound 0308-68 (0.15 g, 0.4 mmol), 25 mL ofdichloromethane and triethylamine (0.162 g, 1.6 mmol). The reactionmixture was stirred for 2 hours at 0° C. The reaction was washed withwater and dried over sodium sulfate, filtered and evaporated to give thetitle product 0310-69 as a brown oil (185 mg, 92%): LCMS: 505 [M+1]⁺.

Step 29b.N¹-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-yl)-N⁶-hydroxyadipamide(Compound 69)

The freshly prepared hydroxylamine solution (6 mL, 4 mmol) was placed in25 mL flask. Compound 0310-69 (185 mg, 0.38 mmol) was added to thissolution and stirred at room temperature for 4 hours. The mixture wasneutralized with acetic acid/methanol. The mixture was concentratedunder reduce pressure. The residue was purified by preparative HPLC togive the title compound 69 as a white solid (150 mg, 74%): LCMS: 506[M+1]⁺; ¹H NMR (DMSO-d₆): 1.58 (m, 4H), 1.98 (t, J=5.7 Hz, 2H), 2.46 (t,2H), 3.30 (s, 3H), 3.78 (t, J=4.2 Hz, 2H), 4.32 (t, J=5.1 Hz, 2H), 7.28(s, 1H), 7.39 (t, J=9 Hz, 1H), 7.79 (m, 1H), 8.11 (m, J=2.7 Hz, 1H),8.50 (s, 1H), 8.64 (s, 1H), 8.75 (s, 1H), 9.25 (s, 1H), 9.76 (s, 1H),10.33 (s, 1H).

Example 30 Preparation ofN¹-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-yl)-N⁸-hydroxyoctanediamide(Compound 70) Step 30a. Methyl8-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-ylamino)-8-oxooctanoate(Compound 0310-70)

Methyl 8-chloro-8-oxooctanoate (0.496 g, 2.4 mmol) was added to asolution of compound 0308-68 (0.219 g, 0.6 mmol), 30 mL ofdichloromethane and triethylamine (0.48 g, 2.4 mmol). The mixture wasstirred for 2 hours at 0° C. The reaction was washed with water anddried over sodium sulfate, filtered and evaporated to give the titleproduct 0310-70 as a brown oil (281 mg, 88%): LCMS: 533 [M+1]⁺. ¹H NMR(DMSO-d₆), 1.35 (m, 4H), 1.58 (m, 2H), 1.61 (m, 2H), 2.29 (t, J=7.2 Hz,2H), 2.41 (t, J=7.2 Hz, 2H), 3.35 (s, 3H), 3.77 (t, J=4.5 Hz, 2H), 4.32(t, J=4.5 Hz, 2H), 7.28 (s, 1H), 7.40 (t, J=9.3 Hz, 1H), 7.78 (m, 1H),8.11 (m, 1H), 8.50 (s, 1H), 8.74 (s, 1H), 9.24 (s, 1H), 9.76 (s, 1H).

Step 30b.N¹-(4-(3-chloro-4-fluorophenylamino)-7-(2-methoxyethoxy)quinazolin-6-yl)-N⁸-hydroxyoctanediamide(Compound 70)

The freshly prepared hydroxylamine solution (6 mL, 4.0 mmol) was placedin 25 mL flask. Compound 0310-70 (281 mg, 0.53 mmol) was added to thissolution and stirred at room temperature for 4 hours. The mixture wasneutralized with acetic acid/methanol. The mixture was concentratedunder reduce pressure. The residue was purified by preparative HPLC togive the title compound 70 as a yellow solid (126 mg, 40%): LCMS: 506[M+1]⁺; ¹H NMR (DMSO-d₆), 1.35 (m, 4H), 1.58 (m, J=6.9 Hz, 2H), 1.61 (m,J=7.2 Hz, 2H), 1.93 (t, J=7.2 Hz, 2H), 2.42 (t, J=7.5 Hz, 2H), 3.35 (s,3H), 3.77 (t, J=4.5 Hz, 2H), 4.32 (t, J=4.5 Hz, 2H), 7.28 (s, 1H), 7.40(t, J=9.3 Hz, 1H), 7.78 (m, 1H), 8.11 (m, J=2.4 Hz, 1H), 8.50 (s, 1H),8.62 (d, J=1.5 Hz, 1H), 8.75 (s, 1H), 9.25 (s, 1H), 9.76 (s, 1H), 10.31(s, 1H).

Example 31 Preparation of7-(4-(3-ethynyl-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 75) Step 31a. 2-Bromo-1-fluoro-4-nitrobenzene (compound 0602)

To a solution of 1-bromo-2-fluorobenzene (35.0 g, 200 mmol) in 200 mL ofconcentrated sulfuric acid was added 20 mL of 68% nitric acid. Thetemperature of the mixture was maintained below 20° C. After theaddition was completed, the mixture was stirred at 10° C. overnight,then diluted with ice water. The resulting solid was collected byfiltration. The solid was recrystallized from petroleum ether to givethe title compound 0602 as a yellow solid (38 g, 89%): m.p. 55.8-56.7°C., ¹H NMR (DMSO-d₆): δ 7.66 (t, J=9 Hz, 1H), 8.32 (m, 1H), 8.58 (dd,J=3 Hz, 6 Hz, 1H).

Step 31b. ((2-Fluoro-5-nitrophenyl)ethynyl)trimethylsilane (Compound0603)

A mixture of compound 0602 (11.0 g, 50 mmol), ethynyltrimethylsilane(7.5 g, 75 mmol), triphenylphosphine (0.5 g) and palladium (II) acetate(0.25 g) in 125 mL of deaerated triethylamine was heated at 100° C.overnight under argon. The reaction was cooled and was filtrated, andthe filtrate was concentrated to a dark brown oil which was distilledunder reduce pressure to give title compound 0603 as a light brown solid(4.7 g, 40%). ¹H NMR (CDCl₃): δ 0.3 (s, 9H, SiCH), 7.22 (t, J=9.0 Hz,1H), 8.2-8.5 (m, 2H).

Step 31c. 4-Fluoro-3-((trimethylsilyl)ethynyl)benzenamine (Compound0604)

In 25 mL of methanol was mixed with compound 0603 (3.5 g, 14.8 mmol) andiron filings (4.14 g, 74.0 mmol). To this mixture was added concentratedhydrochrolic acid and water to adjust pH 4-5. The mixture was heated toreflux for 3 hours, cooled, and filtrated through silica gel. Thefiltrate was concentrated to yield a yellow solid residue which was thenextracted with ether. The combined organic phase was dried overmagnesium sulfate and concentrated to give the title compound 0604 as abrown solid (2.69 g, 88%): LCMS 208 [M+1]⁺.

Step 31d. 3-Ethynyl-4-fluorobenzenamine (Compound 0605)

Compound 0604 obtained above was treated with 100 mg potassium hydroxidein 20 mL of methanol at room temperature overnight. The solution wasconcentrated, dilute with water, brought to neutrality, and thenextracted with ether. The combined organic phase was dried overmagnesium sulfate, concentrated to yield the title compound 0605 as abrown oil (1.49 g, 85%): LCMS 136 [M+1]⁺. The product was used in thenext step without further purification.

Step 31e. 4-(3-Ethynyl-4-fluorophenylamino)-7-methoxyquinazolin-6-ylacetate (Compound 0606)

A mixture of 4-chloro-7-methoxyquinazolin-6-yl acetate (compound 0105)(252 mg, 1.0 mmol) and 3-ethynyl-4-fluorobenzenamine (605) (200 mg, 1.5mmol) in isopropanol (10 mL) was stirred and heated to reflux for 3hours. The mixture was cooled to room temperature and resultingprecipitate was isolated. The solid was then dried to give the titlecompound 0606 (260 mg, 74.0%) as a light yellow solid: LCMS: 352 [M+1]⁺.

Step 31f. 4-(3-Ethynyl-4-fluorophenylamino)-7-methoxyquinazolin-6-ol(Compound 0607)

A mixture of compound 0606 (260 mg, 0.74 mmol), LiOH H₂O (250 mg, 5.8mmol) in methanol (25 ml) and H₂O (25 ml) was stirred at roomtemperature for 0.5 hour. The mixture was neutralized by addition ofdilution acetic acid. The precipitate was isolated and dried to give thetitle compound 0607 (234 mg, 100%) as a grey solid: LCMS: 310 [M+1]⁺.

Step 31g. Ethyl7-(4-(3-ethynyl-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0608-75)

The title compound 0608-75 was prepared as a yellow solid (300 mg,87.0%) from compound 607 (230 mg, 0.74 mmol) and ethyl 7-bromoheptanoate(176 mg, 0.74 mmol) using a procedure similar to that described forcompound 0110-1 (Example 1): LCMS: 466 [M+1]⁺.

Step 31h.7-(4-(3-Ethynyl-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 75)

The title compound 75 was prepared as a white solid (176 mg, 70%) fromcompound 0608 (250 mg, 0.54 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): mp 150.4˜164.5° C. (dec); LCMS:453 [M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.33 (m, 2H), 1.48 (m, 4H), 1.80 (m,2H), 1.94 (t, J=7.2 Hz, 2H), 3.91 (s, 3H), 4.10 (t, J=6.0 Hz, 2H), 4.51(s, 1H), 7.17 (s, 1H), 7.31 (t, J=7.5 Hz, 1H), 7.77 (s, 1H), 7.85 (m,1H), 7.98 (m, 1H), 8.45 (s, 1H), 8.65 (s, 1H), 9.47 (s, 1H), 10.33 (s,1H).

Example 32 Preparation of(R)—N-hydroxy-6-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)-hexanamide(Compound 77) Step 32a.(R)-7-Methoxy-4-(1-phenylethylamino)quinazolin-6-ol (Compound 0701-77)

A mixture of compound 0105 (2.0 g, 8.0 mmol), (R)-1-phenylethanamine(2.91 g, 24.0 mmol) and isopropanol (50 mL) was stirred at 60° C.overnight. Iospropanol was removed and the residue was purified bycolumn chromatography to give the title compound 0701-77 (1.32 g, 56%).LCMS: 296 [M+1]⁺.

Step 32b. (R)-Ethyl6-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)hexanoate(Compound 0702-77)

A mixture of compound 0701-77 (500.0 mg, 1.69 mmol), K₂CO₃ (700.0 mg,5.07 mmol), ethyl 6-bromohexanoate (378.0 mg, 1.69 mmol) and DMF (20 mL)was heated at 60° C. for 3 h. The DMF was moved under reduced pressure,the residue was suspended in water, and the resulting solid wascollected and dried to give the title compound 0702-77 (320 mg, 43%).LCMS: 438 [M+1]⁺.

Step 32c.(R)—N-Hydroxy-6-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)-hexanamide(Compound 77)

A mixture of compound 0702-77 (320.0 mg, 0.73 mmol) and 1.77 mol/LNH₂OH/MeOH (4.0 mL, 6.77 mmol) was stirred at room temperature for 0.5h. The reaction mixture was neutralized with AcOH and concentrated. Theresidue was suspended in water and the resulting solid was isolated anddried to give crude product. This crude product was purified by pre-HPLCto give the title compound 77 (36 mg, 12%). LCMS: 425 [M+1]⁺; ¹H NMR(DMSO-d₆): δ 1.46 (m, 2H), 1.59 (m, 5H), 1.82 (m, 2H), 2.01 (t, J=8.7Hz, 2H), 3.90 (s, 3H), 4.10 (t, J=6.3 Hz, 2H), 5.63 (m, 1H), 7.09 (s,1H), 7.21 (m, 1H), 7.32 (m, 2H), 7.42 (d, J=7.2 Hz, 2H), 7.75 (s, 1H),8.06 (d, J=8.4 Hz, 1H), 8.27 (s, 1H), 8.67 (s, 1H), 10.36 (s, 1H).

Example 33 Preparation of(R)—N-hydroxy-6-(4-(1-phenylethylamino)-quinazolin-6-yloxy)hexanamide(Compound 78) Step 33a. (R)-4-(1-Phenylethylamino)quinazolin-6-ol(Compound 0701-78)

A mixture of compound 0204 (1.0 g, 4.5 mmol) and(R)-1-(3-chloro-4-fluoro-phenyl)ethanamine (0.87 g, 5.0 mmol) inisopropanol (45 mL) was stirred at 90° C. for 1 hour. The mixture wascooled to room temperature and the resulting precipitate was isolated.The solid was washed in turn with isopropanol and methanol, dried toprovide the title compound (R)-4-(1-phenylethylamino)quinazolin-6-ylacetate as a yellow solid (0.62 g, 61%): LCMS 308 [M+1]⁺.

A mixture of the above product (0.7 g, 2.3 mmol) and lithium hydroxidemonohydrate (0.29 g, 6.81 mmol) in methanol (10 mL)/water (15 mL) wasstirred at room temperature for 1 hour. The pH was adjusted to 4 withacetic acid and filtered. The collected yellow solid was washed by waterand dried to obtained title compound 0701-78 as a yellow solid (0.42 g,62%), LCMS 266 [M+1]⁺.

Step 33b. (R)-Ethyl6-(4-(1-phenylethylamino)quinazolin-6-yloxy)hexanoate (Compound 0702-78)

A mixture of compound 0701-78 (0.31 g, 1.2 mmol), ethyl 6-bromohexanoate(0.27 g, 1.2 mmol) and K₂CO₃ (0.8 g, 5.8 mmol) in DMF (15 mL) wasstirred and heated to 80° C. for 2 hours. The mixture was filtered andthe filtrate was evaporated. The resulting solid was washed with etherto obtain the title compound 0702-78 as a pale yellow solid (0.2 g,42.5%), LCMS 408 [M+1]⁺.

Step 33c.(R)—N-Hydroxy-6-(4-(1-phenylethylamino)quinazolin-6-yloxy)hexanamide(Compound 78)

The title compound 78 was prepared as a pale yellow solid (42 mg, 26%)from compound 0702-78 (168 mg, 0.41 mmol) using a procedure similar tothat described for compound 77 (Example 32): LCMS 395 [M+1]⁺, ¹H NMR(DMSO-d₆): δ 1.47 (m, 2H), 1.52 (m, 2H) 1.65 (d, J=7.2 Hz, 3H) 1.71 (m,2H), 2.05 (t, J=3.9 Hz, 2H), 4.04 (t, J=6.3 Hz, 2H), 5.56 (q, J=6.3 Hz,1H) 7.13 (t, J=7.2 Hz, 1H), 7.26 (t, J=7.8 Hz, 2H), 7.32 (dd, J=2.7,J=9.0 Hz, 1H) 7.39 (d, J=7.2 Hz, 2H), 7.56 (d, J=7.2 Hz, 1H), 7.65 (m,1H), 8.26 (s, 1H).

Example 34 Preparation of(R)—N-hydroxy-7-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)heptanamide(Compound 79) Step 34a. (R)-Ethyl7-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)heptanoate(Compound 0702-79)

A mixture of compound 0701-79 (500 mg, 1.69 mmol), K₂CO₃ (700 mg, 5.07mmol), ethyl 7-bromoheptanoate (401 mg, 1.69 mmol) and DMF (20 mL) washeated at 60° C. for 3 h. The DMF was removed under reduced pressure andthe residue was suspended in water. The resulting solid was collectedand dried to give the title compound 0702-79 (340 mg, 44%). LCMS: 452[M+1]⁺.

Step 34b.(R)—N-Hydroxy-7-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)heptanamide(Compound 79)

The title compound 79 was prepared (41 mg, 12%) from compound 0702-79(340 mg, 0.75 mmol) using a procedure similar to that described forcompound 77 (Example 32): LCMS: 439 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.34 (m,2H), 1.52 (m, 4H), 1.58 (d, J=7.5 Hz, 2H), 1.80 (m, 2H), 1.99 (t, J=8.7Hz, 2H), 3.89 (s, 3H), 4.10 (t, J=6.3 Hz, 2H), 5.62 (m, 1H), 7.08 (s,1H), 7.20 (m, 1H), 7.31 (m, 2H), 7.41 (d, J=7.2 Hz, 2H), 7.74 (s, 1H),8.05 (d, J=8.1 Hz, 1H), 8.26 (s, 1H), 8.63 (s, 1H), 10.32 (s, 1H).

Example 35 Preparation of(S)—N-hydroxy-7-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)heptanamide(Compound 80) Step 35a.(S)-7-Methoxy-4-(1-phenylethylamino)quinazolin-6-ol (Compound 0701-80)

The title compound 0701-80 was prepared as a yellow solid (556 mg,62.8%) from compound 0105 (750 mg, 3.0 mmol) and (S)-1-phenylethanamine(1089 mg, 9.0 mmol) using a procedure similar to that described forcompound 0701-77 (Example 32): LCMS: 296 [M+1]⁺.

Step 35b. (S)-Ethyl7-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)heptanoate(Compound 0702-80)

The title compound 0702-80 was prepared as a yellow solid (160 mg,70.95%) from compound 701-80 (148 mg, 0.5 mmol) and ethyl7-bromoheptanoate (120 mg, 0.5 mmol) using a procedure similar to thatdescribed for compound 0702-77 (Example 32): LCMS: 452 [M+1]⁺.

Step 35c.(S)—N-hydroxy-7-(7-methoxy-4-(1-phenylethylamino)quinazolin-6-yloxy)heptanamide(Compound 80)

The title compound 80 was prepared as a white solid (95 mg, 61.9%) fromcompound 0702-80 (160 mg, 0.35 mmol) and fresh NH₂OH/CH₃OH (3 mL, 5.31mmol) using a procedure similar to that described for compound 77(Example 32): m.p. 106.7˜111.3° C., LCMS: 439 [M+1]⁺, ¹H NMR (DMSO-d₆):δ 1.42 (m, 6H), 1.57 (d, J=6.6 Hz, 3H), 1.79 (m, 2H), 1.95 (t, J=7.2 Hz,2H), 3.88 (s, 3H), 4.08 (t, J=6.9 Hz, 2H), 5.62 (m, J=6.6 Hz, 1H), 7.06(s, 1H), 7.21 (t, J=7.5 Hz, 1H), 7.30 (t, J=7.5 Hz, 2H), 7.41 (d, J=7.5Hz, 2H), 7.75 (s, 1H), 8.15 (d, J=9.6 Hz, 1H), 8.29 (s, 1H), 8.60 (s,1H), 10.30 (s, 1H).

Example 36 Preparation of(R)-7-(4-(1-(4-fluorophenyl)ethylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 81) Step 36a.(R)-4-(1-(4-Fluorophenyl)ethylamino)-7-methoxyquinazolin-6-ol (Compound0701-81)

The title compound 0701-81 was prepared as a yellow solid (495 mg,52.71%) from compound 0105 (750 mg, 3.0 mmol) and(R)-1-(4-fluorophenyl)ethanamine (1251 mg, 9.0 mmol) using a proceduresimilar to that described for compound 0701-77 (Example 32): LCMS: 314[M+1]⁺.

Step 36b. (R)-Ethyl7-(4-(1-(4-fluorophenyl)ethylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0702-81)

The title compound 0702-81 was prepared as a yellow solid (190 mg,81.0%) from compound 0701-81 (156 mg, 0.5 mmol) and ethyl7-bromoheptanoate (120 mg, 0.5 mmol) using a procedure similar to thatdescribed for compound 0702-77 (Example 32): LCMS: 470 [M+1]⁺.

Step 36c.(R)-7-(4-(1-(4-Fluorophenyl)ethylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 81)

The title compound 81 was prepared as a white solid (100 mg, 54.12%)from compound 0702-81 (190 mg, 0.40 mmol) and fresh NH₂OH/CH₃OH (3 mL,5.31 mmol) using a procedure similar to that described for compound 77(Example 32): m.p. 118.2-144.3° C., LCMS: 457 [M+1]⁺, ¹H NMR (DMSO-d₆):δ 1.33 (m, 2H), 1.47 (m, 4H), 1.56 (d, J=7.2 Hz, 3H), 1.78 (m, 2H), 1.95(t, J=7.2 Hz, 2H), 3.87 (s, 1H), 4.07 (t, J=6.0 Hz, 2H), 5.60 (m, 1H),7.06 (s, 1H), 7.11 (t, J=9.0 Hz, 2H), 7.44 (m, 2H), 7.71 (s, 1H), 8.04(d, J=7.5 Hz, 1H), 8.25 (s, 1H), 8.65 (s, 1H), 10.33 (s, 1H).

Example 37 Preparation of(R)-7-(4-(1-(4-chlorophenyl)ethylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 82) Step 37a.(R)-4-(1-(4-Chlorophenyl)ethylamino)-7-methoxyquinazolin-6-ol (Compound0701-82)

The title compound 0701-82 was prepared as a yellow solid (0.65 g, 49%)from compound 0105 (1.0 g, 4 mmol) and (R)-1-(4-chlorophenyl)ethanamine(1.87 g, 12 mmol) using a procedure similar to that described forcompound 0701-77 (Example 32): LCMS: 300 [M+1]⁺.

Step 37b. (R)-Ethyl7-(4-(1-(4-chlorophenyl)ethylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0702-82)

The title compound 0702-82 was prepared as a yellow solid (460 mg, 56%)from compound 0701-82 (550 mg, 1.7 mmol) and ethyl 7-bromoheptanoate(404 mg, 1.7 mmol) using a procedure similar to that described forcompound 0702-77 (Example 32): LCMS: 486 [M+1]⁺.

Step 37c.(R)-7-(4-(1-(4-Chlorophenyl)ethylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 82)

The title compound 82 was prepared as a white solid (145 mg, 29%) fromcompound 0702-81 510 mg, 1.05 mmol) and fresh 0.77 mol/L NH₂OH/MeOH (4.7mL, 8.4 mmol) using a procedure similar to that described for compound77 (Example 32): LCMS: 473 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.34 (m, 2H),1.47 (m, 4H), 1.57 (d, J=6.9 Hz, 3H), 1.80 (m, 2H), 1.97 (t, J=7.2 Hz,2H), 3.89 (s, 3H), 4.10 (t, J=6.6 Hz, 2H), 5.57 (m, 1H), 7.08 (s, 1H),7.38 (d, J=8.4 Hz, 2H), 7.43 (d, J=8.4 Hz, 2H), 7.73 (s, 1H), 8.04 (d,J=7.8 Hz, 1H), 8.23 (s, 1H), 8.64 (s, 1H), 10.33 (s, 1H).

Example 38 Preparation of(R)—N-hydroxy-7-(7-methoxy-4-(1-(4-methoxyphenyl)ethylamino)quinazolin-6-yloxy)-heptanamide(Compound 83) Step 38a.(R)-7-Methoxy-4-(1-(4-methoxyphenyl)ethylamino)quinazolin-6-ol (Compound0701-83)

A mixture of compound 0105 (1.0 g, 4.0 mmol),(R)-1-(4-methoxyphenyl)ethanamine (1.81 g, 12.0 mmol) and isopropanol(25 mL) was stirred at 60° C. overnight. Iospropanol was removed and theresidue was purified by column chromatogram to give the title compound0701-83 (0.81 g, 62%). LCMS: 326 [M+1]⁺.

Step 38b. (R)-Ethyl7-(7-methoxy-4-(1-(4-methoxyphenyl)ethylamino)quinazolin-6-yloxy)heptanoate(Compound 0702-83)

A mixture of compound 0701-83 (630 mg, 1.94 mmol), K₂CO₃ (804 mg, 5.8mmol), ethyl 7-bromoheptanoate (459 mg, 1.94 mmol) and DMF (20 mL) washeated to 60° C. for 3 h. The DMF was moved away under reduced pressure,the residue was suspended in water, and the solid was collected anddried to give the title compound 0703-83 (440 mg, 47%). LCMS: 482[M+1]⁺.

Step 38c.(R)—N-Hydroxy-7-(7-methoxy-4-(1-(4-methoxyphenyl)ethylamino)-quinazolin-6-yloxy)-heptanamide(Compound 83)

A mixture of compound 0702-83 (530 mg, 1.1 mmol) and 1.77 mol/LNH₂OH/MeOH (5 mL, 8.8 mmol) was stirred at room temperature for 0.5 h.The reaction mixture was neutralized with AcOH and then the mixture wasconcentrated and the residue was suspended in water, the precipitate wasisolated and dried to give crude product. This product was purified bypre-HPLC to give the title compound 83 (151 mg, 29%). LCMS: 469 [M+1]⁺;¹H NMR (DMSO-d₆): δ 1.32 (m, 2H), 1.45 (m, 4H), 1.54 (d, J=6.9 Hz, 3H),1.78 (m, 2H), 1.95 (t, J=7.2 Hz, 2H), 3.69 (s, 3H), 3.87 (s, 3H), 4.07(t, J=6.3 Hz, 2H), 5.56 (m, 1H), 6.87 (d, J=8.7 Hz, 2H), 7.05 (s, 1H),7.31 (d, J=8.7 Hz, 2H), 7.70 (s, 1H), 7.96 (d, J=8.1 Hz, 1H), 8.26 (s,1H), 8.62 (s, 1H), 10.31 (s, 1H).

Example 39 Preparation of7-(4-(Benzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 85) Step 39a. 4-(Benzylamino)-7-methoxyquinazolin-6-ol(0701-85)

Benzylamine (1.28 g, 12.0 mmol) was added into a mixture of compound0105 (1.0 g, 4.0 mmol) and 2-propanol (50 ml). The reaction mixture wasthen stirred at reflux for 3 hours. The mixture was cooled to roomtemperature and the resulting precipitate was isolated. The solid wasthen dried to give the title compound 0701-85 as a yellow solid (854 mg,76%): LCMS: 282 [M+1]⁺.

Step 39b. Ethyl7-(4-(benzylamino)-7-methoxyquinazolin-6-yloxy)heptanoate (Compound0702-85)

The title compound 0702-85 was prepared as a yellow solid liquid (270mg, 62%) from compound 0701-85 (281 mg, 1.0 mmol) and ethyl7-bromoheptanoate (236 mg, 1 mmol) using a procedure similar to thatdescribed for compound 0702-77 (Example 32): LCMS: 438 [M+1]⁺.

Step 39c.7-(4-(Benzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 85)

The title compound 85 was prepared as a yellow solid (64 mg, 24%) fromcompound 0702-85 (270 mg, 0.62 mmol) using a procedure similar to thatdescribed for compound 77 (Example 32): LCMS: 425 [M+1]⁺; ¹H NMR(DMSO-d₆): δ 1.32 (m, 2H), 1.42 (m, 2H), 1.51 (m, 2H), 1.76 (m, 2H),1.94 (t, J=7.2 Hz, 2H), 3.88 (s, 3H), 4.03 (t, J=6.3 Hz, 2H), 4.76 (d,J=5.4 Hz, 2H), 7.08 (s, 1H), 7.21 (t, J=6.0 Hz, 2H), 7.30 (t, J=6.0 Hz,2H), 7.33 (t, J=6.6 Hz, 1H), 7.63 (s, 1H), 8.29 (s, 1H), 8.42 (t, J=6.0Hz, 1H), 8.64 (s, 1H), 10.32 (s, 1H).

Example 40 Preparation of7-(4-(4-fluorobenzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 86) Step 40a. 4-(4-Fluorobenzylamino)-7-methoxyquinazolin-6-ol(Compound 0701-86)

The title compound 0701-86 was prepared as a yellow solid (489 mg,54.5%) from compound 0105 (750 mg, 3.0 mmol) and(4-fluorophenyl)methanamine (1125 mg, 9.0 mmol) using a proceduresimilar to that described for compound 0701-77 (Example 32): LCMS: 300[M+1]⁺.

Step 40b. Ethyl7-(4-(4-fluorobenzylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0702-86)

The title compound 0702-86 was prepared as a yellow liquid (408 mg,89.67%) from compound 0701-86 (300 mg, 1.0 mmol), ethyl7-bromoheptanoate (237 mg, 1.0 mmol) using a procedure similar to thatdescribed for compound 0702-77 (Example 32): LCMS: 456 [M+1]⁺.

Step 40c.7-(4-(4-fluorobenzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 86)

The title compound 86 was prepared as a white solid (300 mg, 69.97%)from compound 0702-86 (442 mg, 0.97 mmol) and fresh NH₂OH/CH₃OH (4 mL,7.08 mmol) using a procedure similar to that described for compound 77(Example 32): LCMS: 443 [M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.31˜1.54 (m, 6H),1.77 (m, 2H), 1.94 (t, J=7.5 Hz, 2H), 3.88 (s, 3H), 4.03 (t, J=6.3 Hz,2H), 4.74 (d, J=5.4 Hz, 2H), 7.11 (m, 3H), 7.38 (m, 2H), 7.68 (s, 1H),8.30 (s, 1H), 8.40 (m, 1H), 8.60 (s, 1H), 10.30 (s, 1H).

Example 41 Preparation of7-(4-(3,4-difluorobenzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 87) Step 41a.4-(3,4-Difluorobenzylamino)-7-methoxyquinazolin-6-ol (Compound 0701-87)

The title compound 0701-87 was prepared as a light yellow solid (500 mg,52.6%) from compound 105 (750 mg, 3.0 mmol) and(3,4-difluorophenyl)methanamine (1072 mg, 7.5 mmol) using a proceduresimilar to that described for compound 0701-77 (Example 32): LCMS: 318[M+1]⁺.

Step 41b. Ethyl 7-(4-(3,4-difluorobenzylamino)-7-methoxy-4-a,5-dihydroquinazolin-6-yloxy)heptanoate (Compound 0702-87)

The title compound 0702-87 was prepared as a light yellow solid (205 mg,86.7%) from compound 0701-87 (160 mg, 0.5 mmol), ethyl 7-bromoheptanoate(237 mg, 1.0 mmol) using a procedure similar to that described forcompound 0702-77 (Example 32): LCMS: 474 [M+1]⁺.

Step 41c.7-(4-(3,4-difluorobenzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 87)

The title compound 87 was prepared as a white solid (75 mg, 44.5%) fromcompound 0702-87 (173 mg, 0.366 mmol) and fresh NH₂OH/CH₃OH (2 mL, 3.4mmol) using a procedure similar to that described for compound 77(Example 32): LCMS: 461 [M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.30 (m, 2H), 1.50(m, 4H), 1.77 (m, 2H), 1.94 (t, J=7.2 Hz, 2H), 3.88 (s, 1H), 4.03 (t,J=6.6 Hz, 2H), 4.72 (d, J=6.0 Hz, 2H), 7.08 (s, 1H), 7.19 (s, 1H), 7.35(m, 2H), 7.61 (s, 1H), 8.30 (s, 1H), 8.46 (t, J=6.0 Hz, 1H), 8.64 (s,1H), 10.32 (s, 1H).

Example 42 Preparation of7-(4-(3-chloro-4-fluorobenzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 88) Step 42a.4-(3-Chloro-4-fluorobenzylamino)-7-methoxyquinazolin-6-ol (Compound0701-88)

The title compound 0701-88 was prepared as a light yellow solid (500 mg,50.1%) from compound 0105 (750 mg, 3.0 mmol) and(3-chloro-4-fluorophenyl)methanamine (1435 mg, 9 mmol) using a proceduresimilar to that described for compound 0701-77 (Example 32): LCMS: 334[M+1]⁺.

Step 42b. Ethyl7-(4-(3-chloro-4-fluorobenzylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0702-88)

The title compound 0702-88 was prepared as a yellow solid (306 mg,92.02%) from compound 0701-88 (227 mg, 0.68 mmol), ethyl7-bromoheptanoate (161 mg, 0.68 mmol) using a procedure similar to thatdescribed for compound 0702-77 (Example 32): LCMS: 490 [M+1]⁺.

Step 42c.7-(4-(3-Chloro-4-fluorobenzylamino)-7-methoxyquinazolin-6-yloxy)-hydroxyheptanamide(Compound 88)

The title compound 88 was prepared as a white solid (210 mg, 70.02%)from compound 0702-88 (306 mg, 0.63 mmol) and fresh NH₂OH/CH₃OH (3 mL,5.31 mol) using a procedure similar to that described for compound 77(Example 32): m.p. 143.1° C. (decomp.), LCMS: 477 [M+1]⁺, ¹H NMR(DMSO-d₆): δ 1.31 (m, 2H), 1.48 (m, 4H), 1.77 (m, 2H), 1.94 (t, J=7.2Hz, 2H), 3.89 (s, 3H), 4.04 (t, J=6.6 Hz, 2H), 4.74 (d, J=5.4 Hz, 2H),7.09 (s, 1H), 7.35 (d, J=7.8 Hz, 2H), 7.54 (d, J=8.4 Hz, 1H), 7.63 (s,1H), 8.35 (s, 1H), 8.58 (m, 1H), 8.65 (s, 1H), 10.33 (s, 1H), 11.92 (s,1H).

Example 43 Preparation of7-(4-(3-bromobenzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 89) Step 43a. 4-(3-Bromobenzylamino)-7-methoxyquinazolin-6-ol(Compound 0701-89)

The title compound 0701-89 was prepared as a yellow solid (543 mg,50.2%) from compound 0105 (750 mg, 3.0 mmol) and(3-bromophenyl)methanamine (1674 mg, 9 mmol) using a procedure similarto that described for compound 0701-77 (Example 32): LCMS: 360 [M+1]⁺.

Step 43b. Ethyl7-(4-(3-bromobenzylamino)-7-methoxyquinazolin-6-yloxy)heptanoate(Compound 0702-89)

The title compound 0702-89 was prepared as a yellow solid (230 mg,89.15%) from compound 0701-89 (180 mg, 0.5 mmol), ethyl7-bromoheptanoate (120 mg, 0.5 mmol) using a procedure similar to thatdescribed for compound 0702-77 (Example 32): LCMS: 516 [M+1]⁺.

Step 43c.7-(4-(3-bromobenzylamino)-7-methoxyquinazolin-6-yloxy)-N-hydroxyheptanamide(Compound 89)

The title compound 89 was prepared as a white solid (105 mg, 53.96%)from compound 0702-89 (200 mg, 0.39 mmol) and fresh NH₂OH/CH₃OH (3 mL,5.31 mol) using a procedure similar to that described for compound 77(Example 32): LCMS: 503 [M+1]⁺, ¹H NMR (DMSO-d₆): δ 1.31˜1.56 (m, 6H),1.75 (m, 2H), 1.94 (t, J=7.2 Hz, 2H), 3.88 (s, 3H), 4.06 (t, J=6.6 Hz,2H), 4.75 (d, J=5.7 Hz, 2H), 7.08 (s, 1H), 7.27 (t, J=7.5 Hz, 1H),3.7.33 (m, 2H), 7.42 (s, 1H), 7.61 (s, 1H), 7.93 (s, 1H), 8.30 (s, 1H),8.41 (t, J=6.0 Hz, 1H), 8.60 (s, 1H), 10.29 (s, 1H).

Example 44 Preparation of4-(2-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)ethoxy)-N-hydroxybenzamide(Compound 92) Step 44a. Methyl 4-(2-bromoethoxy)benzoate (Compound0502-92)

A mixture of compound 4-hydroxybenzoic acid methyl ester (457.0 mg, 3.0mmol), K₂CO₃ (828 mg, 6 mmol) and 1,2-dibromoethane (10 mL) was heatedat 130° C. for 8 h. The 1,2-dibromoethane was removed under reducedpressure and the residue was suspended in water. The resultingprecipitate was isolated and dried to give the title compound 0502-92 asa white solid (440 mg, 57%). LCMS: 259 [M+1]⁺.

Step 44b. Methyl4-(2-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)ethoxy)benzoate(Compound 0503-92)

A mixture of compound 109 (384 mg, 1.2 mmol), K₂CO₃ (276 mg, 2 mmol),compound 0502-92 (311 mg, 1.2 mmol) and DMF (10 mL) was heated at 40° C.overnight. The DMF was removed under reduced pressure and the residuewas suspended in water. The precipitate was collected and dried to givethe title compound 0503-92 as a white solid (430 mg, 72%). LCMS: 259[M+1]⁺.

Step 44c.4-(2-(4-(3-Chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)ethoxy)-N-hydroxybenzamide(Compound 92)

A mixture of compound 0502-92 (249 mg, 0.5 mmol) and 1.77 mol/LNH₂OH/MeOH (5 mL, 8.85 mmol) was stirred at room temperature for 0.5 h.The reaction mixture was neutralized with AcOH and the mixture wasconcentrated and the residue was suspended in water. The resultingprecipitate was isolated and dried to give crude product. This crudeproduct was purified by pre-HPLC to give the title compound 92 as awhite solid (80 mg, 32%). LCMS: 439 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 2.07 (s,2H), 3.93 (s, 3H), 4.50 (s, 4H), 7.08 (d, J=8.4 Hz, 2H), 7.22 (s, 2H),7.44 (t, J=9.0 Hz, 1H), 7.76 (m, 3H), 7.89 (s, 1H), 8.12 (m, 1H), 8.51(s, 1H), 8.87 (s, 1H), 9.54 (s, 1H), 11.05 (s, 1H).

Example 45 Preparation of7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-methoxyheptanamide(Compound 95)

A mixture of compound 0802 (544 mg, 1.25 mmol) and Inodomethane (0804)(177 mg, 1.25 mmol) and potassium carbonate (1.0 g, 7.25 mmol) inN,N-dimethylformamide (15 mL) was stirred at room temperature for 12hours. The solvent was removed under reduce pressure and the residue wasdissolved in ethyl acetate (50 mL). The organic layer was washed withsaturation aqueous NaHCO₃ (20 mL) and brine (20 mL). The organic layerwas dried over MgSO₄ and concentrated to give the title compound 95 aspale yellow solid (500 mg, 89%). m.p. 195.81˜97.0° C.; LCMS: 449 [M+1]⁺;¹H NMR (DMSO-d₆): δ 1.35 (m, 2H), 1.50 (m, 4H), 1.80 (m, 2H), 1.94 (t,J=7.2 Hz, 2H), 3.54 (s, 3H), 3.92 (s, 3H), 4.12 (t, J=6.3 Hz, 2H), 4.19(s, 1H), 7.19 (m, 2H), 7.40 (t, J=7.8 Hz, 1H), 7.80 (s, 1H), 7.87 (d,J=9.6 Hz, 1H), 7.97 (s, 1H), 8.48 (s, 1H), 9.45 (s, 1H), 10.92 (s, 1H).

Example 46 Preparation ofN-acetoxy-7-(4-(3-chloro-4-fluorophenylamino)-7-methoxyquinazolin-6-yloxy)heptanamide(Compound 96)

A mixture of compound 0801 (50 mg, 0.108 mmol) and Ac₂O (204 mg, 2.0mmol) and AcOH (2 mL) was stirred at room temperature for 1 h. Thereaction mixture was neutralized with NaHCO₃ saturation solution. Theprecipitate was isolated and dried to give product 96 (42 mg, 77%).LCMS: 505 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.40 (m, 2H), 1.50 (m, 2H), 1.55(m, 2H), 1.80 (m, 2H), 2.09 (s, 3H), 2.12 (m, 2H), 3.94 (s, 3H), 4.13(t, J=6.9 Hz, 2H), 7.20 (s, 1H), 7.43 (t, J=9.0 Hz, 1H), 7.78 (m, 1H),7.84 (s, 1H), 8.12 (m, 1H), 8.49 (s, 1H), 9.67 (s, 1H).

Example 47 Preparation ofN-acetoxy-7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)heptanamide(Compound 97)

The title compound 97 was prepared as a solid (45 mg, 86.0%) fromcompound 0802 (48 mg, 0.11 mmol) and Ac₂O (204 mg, 2 mmol) using aprocedure similar to that described for compound 96 (Example 46): LCMS:476.5 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.40 (m, 2H), 1.46 (m, 2H), 1.58 (m,2H), 1.80 (m, 2H), 2.12 (s, 3H), 2.13 (m, 2H), 3.94 (s, 3H), 4.14 (t,J=6.6 Hz, 2H), 4.19 (s, 1H), 7.20 (d, J=6.3 Hz, 2H), 7.40 (t, J=7.8 Hz,1H), 7.83 (s, 1H), 7.89 (d, J=7.8 Hz, 1H), 7.99 (s, 1H), 8.49 (s, 1H),9.50 (s, 1H), 11.55 (s, 1H).

Example 48 Preparation ofN-(cyclohexanecarbonyloxy)-7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)heptanamide(Compound 98)

Compound 0802 (218 mg, 0.5 mmol) and triethylamine (75 mg, 0.75 mmol)were dissolved in acetone (20 mL) and N,N-dimethylformamide (2 mL). Thereaction mixture was cooled to 0° C. and a solution ofcyclohexanecarbonyl chloride (73 mg, 0.5 mmol) in acetone (5 mL) wasadded into the above solution dropwise. The reaction mixture was allowedto raise to ambient temperature and stirred for 1 hour. The mixture wasconcentrated under reduce pressure and the residue was purified bycolumn chromatography to give the title compound 98 as a yellow solid(50 mg, 18%): LCMS: 545 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.21˜1.63 (m, 15H),1.81 (m, 4H), 2.11 (t, J=7.2 Hz, 2H), 3.92 (s, 3H), 4.12 (t, J=7.2 Hz,2H), 4.17 (s, 1H), 7.19 (m, 2H), 7.39 (t, J=7.8 Hz, 1H), 7.81 (s, 1H),7.88 (d, J=8.4 Hz, 1H), 7.97 (s, 1H), 8.47 (s, 1H), 9.45 (s, 1H), 11.50(s, 1H).

Example 49 Preparation of7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-(isobutyryloxy)heptanamide(Compound 99)

The title compound 99 was prepared as a yellow solid (100 mg, 44.0%)from compound 0802 (195 mg, 0.45 mmol) and isobutyryl chloride (48 mg,0.45 mmol) using a procedure similar to that described for compound 98(Example 48): LCMS: 505 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.10 (d, J=7.2 Hz,6H), 1.39 (m, 2H), 1.47 (m, 2H), 1.56 (m, 2H), 1.81 (m, 2H), 2.11 (t,J=7.5 Hz, 2H), 2.68 (m, J=7.2 Hz, 2H), 3.92 (s, 3H), 4.12 (t, J=6.6 Hz,2H), 4.17 (s, 1H), 7.19 (m, 2H), 7.38 (t, J=7.8 Hz, 1H), 7.82 (s, 1H),7.88 (d, J=8.7 Hz, 1H), 7.97 (s, 1H), 8.47 (s, 1H), 9.50 (s, 1H), 11.55(s, 1H).

Example 50 Preparation of7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)-N-(propionyloxy)heptanamide(Compound 100)

The title compound 100 was prepared as a yellow solid (100 mg, 41.0%)from compound 0802 (218 mg, 0.5 mmol) and propionyl chloride (47 mg, 0.5mmol) using a procedure similar to that described for compound 98(Example 48): LCMS: 491 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.05 (t, J=7.5 Hz,3H), 1.39 (m, 2H), 1.48 (m, 2H), 1.56 (m, 2H), 1.81 (m, 2H), 2.12 (t,J=6.6 Hz, 2H), 2.41 (q, J=7.5 Hz, 2H), 3.92 (s, 3H), 4.12 (t, J=6.6 Hz,2H), 4.18 (s, 1H), 7.19 (m, 2H), 7.38 (t, J=7.8 Hz, 1H), 7.80 (s, 1H),7.88 (d, J=8.1 Hz, 1H), 7.97 (s, 1H), 8.47 (s, 1H), 9.45 (s, 1H), 11.53(s, 1H).

Example 51 Preparation ofN-(benzoyloxy)-7-(4-(3-ethynylphenylamino)-7-methoxyquinazolin-6-yloxy)heptanamide(Compound 101)

The title compound 101 was prepared as a yellow solid (150 mg, 56.0%)from compound 0802 (218 mg, 0.5 mmol) and benzoyl chloride (72 mg, 0.5mmol) using a procedure similar to that described for compound 98(Example 48): LCMS: 539 [M+1]⁺; ¹H NMR (DMSO-d₆): δ 1.51 (m, 4H), 1.61(m, 2H), 1.84 (m, 2H), 2.21 (t, J=7.5 Hz, 2H), 3.93 (s, 3H), 4.14 (t,J=6.9 Hz, 2H), 4.19 (s, 1H), 7.19 (m, 2H), 7.38 (t, J=7.8 Hz, 1H), 7.55(m, 2H), 7.72 (t, J=7.8 Hz, 1H), 7.82 (s, 1H), 7.89 (d, J=8.7 Hz, 1H),7.99 (m, 3H), 8.48 (s, 1H), 9.48 (s, 1H), 11.88 (s, 1H).

Biological Assays:

As stated hereinbefore the derivatives defined in the present inventionpossess anti-proliferation activity. These properties may be assessed,for example, using one or more of the procedures set out below:

(a) An In Vitro Assay which Determines the Ability of a Test Compound toInhibit EGFR Kinase.

The ability of compounds to inhibit receptor kinase (EGFR) activity wasassayed using HTScan™ EGF Receptor Kinase Assay Kits (Cell SignalingTechnologies, Danvers, Mass.). EGFR tyrosine kinase was obtained asGST-kinase fusion protein which was produced using a baculovirusexpression system with a construct expressing human EGFR(His672-Ala1210) (GenBank Accession No. NM_(—)005228) with anamino-terminal GST tag. The protein was purified by one-step affinitychromatography using glutathione-agarose. An anti-phosphotyrosinemonoclonal antibody, P-Tyr-100, was used to detect phosphorylation ofbiotinylated substrate peptides (EGFR, Biotin-PTP1B (Tyr66). Enzymaticactivity was tested in 60 mM HEPES, 5 mM MgCl₂ 5 mM MnCl₂ 200 μM ATP,1.25 mM DTT, 3 μM Na₃VO₄, 1.5 mM peptide, and 50 ng EGF Receptor Kinase.Bound antibody was detected using the DELFIA system (PerkinElmer,Wellesley, Mass.) consisting of DELFIA® Europium-labeled Anti-mouse IgG(PerkinElmer, #AD0124), DELFIA® Enhancement Solution (PerkinElmer,#1244-105), and a DELFIA® Streptavidin coated, 96-well Plate(PerkinElmer, AAAND-0005). Fluorescence was measured on a WALLAC Victor2 plate reader and reported as relative fluorescence units (RFU). Datawere plotted using GraphPad Prism (v4.0a) and IC50's calculated using asigmoidal dose response curve fitting algorithm.

Test compounds were dissolved in dimethylsulphoxide (DMSO) to give a 20mM working stock concentration. Each assay was setup as follows: Added100 μl of 10 mM ATP to 1.25 ml 6 mM substrate peptide. Diluted themixture with dH₂0 to 2.5 ml to make 2×ATP/substrate cocktail ([ATP]=400mM, [substrate]=3 mM). Immediately transfer enzyme from −80° C. to ice.Allowed enzyme to thaw on ice. Microcentrifuged briefly at 4° C. tobring liquid to the bottom of the vial. Returned immediately to ice.Added 10 μl of DTT (1.25 mM) to 2.5 ml of 4×HTScan™ Tyrosine KinaseBuffer (240 mM HEPES pH 7.5, 20 mM MgCl₂, 20 mM MnCl, 12 mM NaVO₃) tomake DTT/Kinase buffer. Transfer 1.25 ml of DTT/Kinase buffer to enzymetube to make 4× reaction cocktail ([enzyme]=4 ng/μL in 4× reactioncocktail). Incubated 12.5 μl of the 4× reaction cocktail with 12.5μl/well of prediluted compound of interest (usually around 10 μM) for 5minutes at room temperature. Added 25 μl of 2×ATP/substrate cocktail to25 μl/well preincubated reaction cocktail/compound. Incubated reactionplate at room temperature for 30 minutes. Added 50 μl/well Stop Buffer(50 mM EDTA, pH 8) to stop the reaction. Transferred 25 μl of eachreaction and 75 μl dH₂O/well to a 96-well streptavidin-coated plate andincubated at room temperature for 60 minutes. Washed three times with200 μl/well PBS/T (PBS, 0.05% Tween-20). Diluted primary antibody,Phospho-Tyrosine mAb (P-Tyr-100), 1:1000 in PBS/T with 1% bovine serumalbumin (BSA). Added 100 μl/well primary antibody. Incubated at roomtemperature for 60 minutes. Washed three times with 200 μl/well PBS/T.Diluted Europium labeled anti-mouse IgG 1:500 in PBS/T with 1% BSA.Added 100 μl/well diluted antibody. Incubated at room temperature for 30minutes. Washed five times with 200 μl/well PBS/T. Added 100 μl/wellDELFIA® Enhancement Solution. Incubated at room temperature for 5minutes. Detected 615 nm fluorescence emission with appropriateTime-Resolved Plate Reader.

(b) An In Vitro Assay which Determines the Ability of a Test Compound toInhibit the EGF-Stimulated EGFR Phosphorylation.

Allowed A431 cell growth in a T75 flask using standard tissue cultureprocedures until cells reach near confluency (˜1.5×10⁷) cells; D-MEM,10% FBS). Under sterile conditions dispensed 100 μl of the cellsuspension per well in 96-well microplates (x cells plated per well).Incubated cells and monitor cell density until confluency is achievedwith well-to-well consistency; approximately three days. Removedcomplete media from plate wells by aspiration or manual displacement.Replaced media with 50 μl of pre-warmed serum free media per well andincubated 4 to 16 hours. Made two fold serial dilutions of inhibitorusing pre-warmed D-MEM so that the final concentration of inhibitorrange from 10 μM to 90 μM. Removed media from A431 cell plate. Added 100μl of serial diluted inhibitor into cells and incubate 1 to 2 hours.Removed inhibitor from plate wells by aspiration or manual displacement.Added either serum free media for resting cells (mock) or serum freemedia with 100 ng/ml EGF. Used 100 μl of resting/activation media perwell. Allowed incubation at 37° C. for 7.5 minutes. Removed activationor stimulation media manually or by aspiration. Immediately fixed cellswith 4% formaldehyde in 1×PBS. Allowed incubation on bench top for 20minutes at RT with no shaking. Washed five times with 1×PBS containing0.1% Triton X-100 for 5 minutes per Wash. Removed Fixing Solution. Usinga multi-channel pipettor, added 200 μl of Triton Washing Solution(1×PBS+0.1% Triton X-100). Allowed wash to shake on a rotator for 5minutes at room temperature. Repeated washing steps 4 more times afterremoving wash manually. Using a multi-channel pipettor, blockedcells/wells by adding 100 μl of LI-COR Odyssey Blocking Buffer to eachwell. Allowed blocking for 90 minutes at RT with moderate shaking on arotator. Added the two primary antibodies into a tube containing OdysseyBlocking Buffer. Mixed the primary antibody solution well beforeaddition to wells (Phospho-EGFR Tyr1045, (Rabbit; 1:100 dilution; CellSignaling Technology, 2237; Total EGFR, Mouse; 1:500 dilution; BiosourceInternational, AHR5062). Removed blocking buffer from the blocking stepand added 40 μl of the desired primary antibody or antibodies in OdysseyBlocking Buffer to cover the bottom of each well. Added 100 μl ofOdyssey Blocking Buffer only to control wells. Incubated with primaryantibody overnight with gentle shaking at RT. Washed the plate fivetimes with 1×PBS+0.1% Tween-20 for 5 minutes at RT with gentle shaking,using a generous amount of buffer. Using a multi-channel pipettor added200 μl of Tween Washing Solution. Allowed wash to shake on a rotator for5 minutes at RT. Repeated washing steps 4 more times. Diluted thefluorescently labeled secondary antibody in Odyssey Blocking Buffer(Goat anti-mouse IRDye™ 680 (1:200 dilution; LI-COR Cat. #926-32220)Goat anti-rabbit IRDye™ 800CW (1:800 dilution; LI-COR Cat. #926-32211).Mixed the antibody solutions well and added 40 μl of the secondaryantibody solution to each well. Incubated for 60 minutes with gentleshaking at RT. Protected plate from light during incubation. Washed theplate five times with 1×PBS+0.1% Tween-20 for 5 minutes at RT withgentle shaking, using a generous amount of buffer. Using a multi-channelpipettor added 200 μl of Tween Washing Solution. Allowed wash to shakeon a rotator for 5 minutes at RT. Repeated washing steps 4 more times.After final wash, removed wash solution completely from wells. Turnedthe plate upside down and tap or blot gently on paper towels to removetraces of wash buffer. Scanned the plate with detection in both the 700and 800 channels using the Odyssey Infrared Imaging System (700 nmdetection for IRDye™ 680 antibody and 800 nm detection for IRDye™ 800CWantibody). Determined the ratio of total to phosphorylated protein(700/800) using Odyssey software and plot the results in Graphpad Prism(V4.0a). Data were plotted using GraphPad Prism (v4.0a) and IC50'scalculated using a sigmoidal dose response curve fitting algorithm.

(c) An In Vitro Assay which Determines the Ability of a Test Compound toInhibit HDAC Enzymatic Activity.

HDAC inhibitors were screened using an HDAC fluorimetric assay kit(AK-500, Biomol, Plymouth Meeting, Pa.). Test compounds were dissolvedin dimethylsulphoxide (DMSO) to give a 20 mM working stockconcentration. Fluorescence was measured on a WALLAC Victor 2 platereader and reported as relative fluorescence units (RFU). Data wereplotted using GraphPad Prism (v4.0a) and IC50's calculated using asigmoidal dose response curve fitting algorithm. Each assay was setup asfollows: Defrosted all kit components and kept on ice until use. DilutedHeLa nuclear extract 1:29 in Assay Buffer (50 mM Tris/Cl, pH 8.0, 137 mMNaCl, 2.7 mM KCl, 1 mM MgCl2). Prepared dilutions of Trichostatin A(TSA, positive control) and tested compounds in assay buffer (5× offinal concentration). Diluted Fluor de Lys™ Substrate in assay buffer to100 uM (50 fold=2× final). Diluted Fluor de Lys™ developer concentrate20-fold (e.g. 50 μl plus 950 μl Assay Buffer) in cold assay buffer.Second, diluted the 0.2 mM Trichostatin A 100-fold in the 1× Developer(e.g. 10 μl in 1 ml; final Trichostatin A concentration in the 1×Developer=2 μM; final concentration after addition to HDAC/Substratereaction=1 μM). Added Assay buffer, diluted trichostatin A or testinhibitor to appropriate wells of the microtiter plate. Added dilutedHeLa extract or other HDAC sample to all wells except for negativecontrols. Allowed diluted Fluor de Lys™ Substrate and the samples in themicrotiter plate to equilibrate to assay temperature (e.g. 25 or 37° C.Initiated HDAC reactions by adding diluted substrate (25 μl) to eachwell and mixing thoroughly. Allowed HDAC reactions to proceed for 1 hourand then stopped them by addition of Fluor de Lys™ Developer (501).Incubated plate at room temperature (25° C.) for 10-15 min. Read samplesin a microtiter-plate reading fluorimeter capable of excitation at awavelength in the range 350-380 nm and detection of emitted light in therange 440-460 nm.

The following TABLE B lists compounds representative of the inventionand their activity in HDAC and EGFR assays. In these assays, thefollowing grading was used: I≧10 μM, 10 μM>II>1 μM, 1 μM>III>0.1 μM, andIV≦0.1 μM for IC₅₀.

TABLE B Compound No. HDAC EGFR HER2/ErB2 VEGFR 1 I IV 2 I IV 3 I IV 4III IV 5 IV IV 6 IV IV IV III 7 I IV IV 8 I IV 9 III IV 10 III IV 11 IVIV III 12 IV IV III 13 I IV 14 II IV 15 IV III 16 III IV 17 IV IV 18 IVIV III 19 I IV 21 II III 22 IV IV 23 IV III 24 IV III 30 IV IV III 36 IVIV 38 II IV 40 IV IV III 42 III IV 43 III IV 44 IV IV III 45 I III 50III III 63 III II 66 III IV 68 II IV 69 III IV 70 IV IV IV 75 IV IV III76 IV IV III 77 IV IV 78 IV III 79 IV IV III I 80 IV II 81 III III 82III III 83 IV I 84 IV III 85 IV IV III II 86 IV III 87 IV III 88 IV IVII 89 IV III 90 IV N/A 91 II IV 92 III IV 93 II IV 94 I IV 102 IV 103 I107 III 112 I 118 II 121 I 124 I 125 III 138 II 139 II 144 III 145 IV IVIV 151 IV IV IV 155 IV IV IV 161 IV III 162 IV IV III 167 IV IV III 168IV IV III 174 I 177 III IV IV 178 III 198 IV IV IV 199 IV IV IV 200 IVIV IV 201 III IV IV 202 III 203 III 204 II 205 II 206 IV IV IV 207 IV IVIV 208 IV 209 IV

A representative number of compounds were assayed against severaldifferent cell lines using the cell proliferation assay:

Cell Proliferation Assay:

Cancer cell lines were plated at 5,000 to 10,000 per well in 96-wellflatted bottomed plates with various concentration of compounds. Thecells were incubated with compounds for 72 hours in the presence of 0.5%of fetal bovine serum. Growth inhibition was accessed by adenosinetriphosphate (ATP) content assay using Perkin Elmer ATPlite kit. ATPliteis an ATP monitoring system based on firefly luciferase. Briefly, 25 μlof mammalian cell lysis solution was added to 50 μl of phenol red-freeculture medium per well to lyse the cells and stabilize the ATP. 25 μlof substrate solution was then added to the well and subsequently theluminescence was measured.

The results are presented below in TABLE C. In these assays, thefollowing grading was used: I≧10 μM, 10 μM>II>1 μM, 1 μM>III>0.1 μM, andIV≦0.1 μM for IC₅₀.

TABLE C Compound No. Cell Line 6 12 18 40 44 66 70 77 79 85 Breast_MCF7IV Breast_MDAMB468 IV Breast_SkBr3 III Colon_HCT116 III III IIIEpidermoid_A431 III Lung_H1703 III III II Lung_H1975 III III IILung_H2122 III III II Lung_H292 IV Lung_H358 III III II Lung_H460 IIIIII II Lung_HCC827 IV III III Pancreas_BxPC3 III IV III II II II II IIII III Pancreas_Capan1 II III II Pancreas_CFPAC III III II I II II II IIPancreas_HPAC II II II II I I I II Pancreas_MiaPaCa2 III III II II IIIII II II Pancreas_PANC1 III III II II II I II II Prostate_22RV1 IIIProstate_PC3 III III

FIG. 1 shows that compounds of the invention, such as compounds 6 and 12are more active than erlotinib and SAHA in EFGR enzyme assay and HDACenzyme assay. In the EFGR enzyme assay, compounds of the invention ismore potent than erlotinib by approximately 15-20 fold. In the HDACenzyme assay, compounds of the invention is more potent than SAHA byapproximately 5-10 fold.

FIG. 2 illustrates the improvement in inhibition of histone acetylationand EGFR phosphorylation by compound 12 as compared with SAHA andErlotinib respectively. Inhibition on both kinase (EGFR) and non-kinase(HDAC) cancer targets by a compound 12.

Table D illustrates the potency of compounds of the invention. Forexample, compound 12 is more active than Erlotinib and SAHA in variouscancer cell lines (IC50 in μM). Cell lines from five major types ofcancer (lung, breast, prostate, colon, and pancreas) responded better tocompound 12 than a combination of erlotinib and SAHA. Surprisingly, thecompounds of the invention are active against cell lines that areresistant to Tarceva® and Iressa®. In these assays, the followinggrading was used: D≧5 μM, 5 μM>C≧0.5 μM, 0.5 μM>B≧0.05 μM, and A≦0.05 μMfor IC₅₀.

TABLE D Erlotinib/SAHA Compound Tumor Line Tumor Type SAHA ErlotinibCombined 12 MDA-MB-231 Breast adenocarcinoma B D B A HCT116 Colon cancerC D C A MCF-7 Breast adenocarcinoma C D C A MDA-MB-468 Breastadenocarcinoma C C C A SKBr3 Breast carcinoma C D C B PC-3 Prostateadenocarcinoma C D C C Caki-1 Renal carcinoma B B B A A431 Epidermoidcarcinoma C C C B 22RV1 Prostate carcinoma B B B B

FIG. 3 shows examples of greater anti-proliferative activity againstseveral different cancer cell lines. FIG. 3 further shows that compoundsof the invention are more potent than SAHA alone, Erlotinib alone, andSAHA and Erlotinib combined.

FIG. 4 displays the potency of compound 12 in induction of apoptosis incolon and breast cancer cells. Compound 12 induced approximately 4-11times more cell apoptosis as measured by increased Caspase 3&7 activity.Erlotinib was inactive at a concentration <20 μM. The high potencydisplayed by compound 12 over Erlotinib suggests that compounds of theinvention can be used to treat tumor cells that are resistant toErlotinib.

FIGS. 5-10 illustrate the efficacy of compound 12 in various tumorxenograft models. Table E below summarizes the in vivo experiments thatwere carried out to give results represented in FIGS. 5-10.

TABLE E Dosing Pre- Method of regimen treatment Model Cancer type Dosagegroups administration (on-off-on) tumor size A431 Epidermoid vehicle IPOnce 156 ± 57 mm³  6 mg/kg daily for 12 mg/kg 21 days 24 mg/kg 48 mg/kgH358 NSCLC vehicle IV - 2 min 7-7-5  84 ± 23 mm³ 15 mg/kg infusion 30mg/kg 60 mg/kg H292 NSCLC vehicle IV - 2 min 7-7-5 116 ± 23 mm³ 15 mg/kginfusion 30 mg/kg 60 mg/kg BxPC3 Pancreatic vehicle IV - 2 min 7-7-2 201± 53 mm³ 10 mg/kg infusion 20 mg/kg 40 mg/kg PC3 Prostate vehicle IV - 2min 7-7-5 195 ± 50 mm³ 10 mg/kg infusion 20 mg/kg 40 mg/kg HCT116 Colonvehicle IV - 2 min 5-2-5  91 ± 23 mm³ 15 mg/kg infusion 30 mg/kg 60mg/kg SAHA 20 mg/kg HCC827 NSCLC vehicle IV - 2 min Once 149 ± 36 mm³(apoptosis/ 30 mg/kg infusion daily for anti- 3 days proliferation)BxPC3 Pancreatic 60 mg/kg IV - 2 min Single IV NA (apoptosis/ infusioninfusion anti- proliferation)A representative protocol for the in vivo experiment is as followed:

1-10×10⁶ human cancer cells were implanted subcutaneously to the athymic(nu/nu) mice. When the tumors reached about 100 mm³ in volume, the micewere treated with the compound by tail vein infusion. Routinely 5 groups(8-12 mice per group) are needed for a typical efficacy study, includingone negative control, one positive control, and three testing groups for3 dose levels of the same compound. Usually a 7-7-5 (on-off-on) regimenwas used for one typical study. The tumor size was measured with anelectronic caliper and body weight measured with a scale twice weekly.The tumors were removed from euthanized mice at the end of the study.One half of each tumor was frozen in dry ice and stored at −80° C. forPK or Western blot analysis. The other half was fixed with formalin. Thefixed tissues were processed, embedded in paraffin and sectioned forimmunohistochemistry staining.

Protocol for Radioisotope Assay for HER2

10 nM HER2 and 0.1 mg/ml polyEY were placed in the reaction buffer and 2mM MnCl₂, 1 μM ATP and 1% DMSO final were added. The reaction mixturewas incubated for 2 hours at room temperature. The conversion rate ofATP was 22%.

HER2 (Accession number: GenBank X03363) is characterized as follows:N-terminal GST-tagged, recombinant, human HER2 amino acids 679-1255,expressed by baculovirus in Sf9 insect cells. Purity >90% by SDS PAGEand Coomassie blue staining. MW=91.6 kDa. Specific Activity of 40 U/mg,where one unit of activity is defined as 1 nmol phosphate incorporatedinto 30 ug/ml Poly (Glu:Tyr) 4:1 substrate per minute at 30° C. with afinal ATP concentration of 100 μM. Enzyme is in 25 mM Tris-HCl, pH 8.0,100 mM NaCl, 0.05% Tween-20, 50% glycerol, 10 mM reduced glutathione,and 3 mM DTT.

REFERENCES

-   1. Meyer, M. et al., EMBO J. 18, 363-374 (1999)-   2. Rahimi, N. et al., J. Biol Chem 275, 16986-16992 (2000)

Compounds of the invention are found to be active against variouskinases. For example, Table F shows inhibition of compound 12 in a panelof kinase assays. Furthermore, Compound 12 is much more active thanErlotinib in Her-2 assay.

TABLE F Assays Concentrations (μM) Inhibition (%) Abl Kinase 5 57 FGFR2Kinase 5 73 FLT-3 Kinase 5 85 VEGFR2 Kinase 5 64 Lck Kinase 5 56 LynKinase 5 95 Ret Kinase 5 93 Her-2 Compound 12 IC50 = 188 nM ErlotinibIC50 = 1473 nM

Example 52 Preparation of Captisol Formulation of Compound 12 A.Preparation of 25, 30, 40, 50 and 60 Mg/Ml Solutions of Compound 12 in30% Captisol (i) With Tartaric Acid

A 30% Captisol formulation was prepared by adding 2.7 ml water to a vialcontaining 0.9 g Captisol. The mixture was then mixed on a vortexer togive ˜3 ml of a clear solution.

In order to prepare a formulation of 25 mg/ml solution of compound 12, 1ml of the 30% Captisol solution was added to a vial containing 25 mg ofcompound 12 and 8.6 mg tartaric acid and the resulting mixture was mixedon a vortexer or sonicated at 30° C. for 15 to 20 minutes to give aclear yellowish solution. The resulting solution is stable at roomtemperature.

In order to prepare a formulation of 30 mg/ml solution of compound 12, 1ml of the 30% Captisol solution was added to a vial containing 30 mg ofcompound 12 and 10.4 mg tartaric acid (1.0 eq) at room temperature.

In order to prepare a formulation of 40 mg/ml solution of compound 12, 1ml of the 30% Captisol solution was added to a vial containing 40 mg ofcompound 12 and 17.9 mg tartaric acid (1.3 eq) at 36° C.

In order to prepare a formulation of 50 mg/ml compound 12 in Captisol, 1ml of 30% Captisol was added to 50 mg compound 12, 22.5 mg tartaric acid(1.3 eq) at 37° C.

In order to prepare a formulation of 60 mg/ml compound 12 in Captisol,the 30% Captisol was added to a vial containing 60 mg compound 12 and26.9 mg tartaric acid (1.3 eq) at 36° C. The solution was diluted in 1×water and 2×D5W. The diluted solution is stable at room temperaturefor >12 h.

(ii) With Citric Acid

In order to prepare a formulation of 25 mg/ml solution of compound 12, 1ml of the 30% Captisol solution was added to a vial containing 25 mg ofcompound 12 and 11.1 mg citric acid (1.0 eq) and the resulting mixturewas mixed on a vortexer or sonicated at room temperature for 15 to 20minutes to give a clear yellowish solution.

(iii) With Hydrochloric Acid

In order to prepare a formulation of 25 mg/ml solution of compound 12, 1ml of the 30% Captisol solution was added to a vial containing 25 mg ofcompound 12 and 57.5 μl hydrochloric acid (1.0 eq) and the resultingmixture was mixed on a vortexer or sonicated at room temperature for 15to 20 minutes to give a clear yellowish solution.

(iv) With Sodium Salt

In order to prepare a formulation of 7.5 mg/ml solution of compound 12,1 ml of the 30% Captisol solution was added to a vial containing 7.5 mgof compound 12 sodium salt and the resulting mixture was mixed on avortexer or sonicated at room temperature for 15 to 20 minutes to give aclear yellowish solution.

B. Filtration of the Solution

The formulations of compound 12 from A (i) was filtered through a 0.2-μmpresterilized filter with >98% recovery.

C. Preparation of a Lyophilisate

The formulations of compound 12 (25 mg/ml) from A (i) and A (iii) werelyophilized to form lyophilisate as a yellow powder.

The lyophilisate resulted from A (i) formulation was chemically stableat following temperatures, −20° C., room temperature, and 40° C. for atleast 2 weeks. It can be stored at 4° C. for greater than 2 weekswithout decomposition. The lyophilisate resulted from A (iii) was stableat −20° C. for at least two weeks.

D. Dilution Study

The formulations of compound 12 from A (i) were diluted with D5W (10-,20-, and 50-fold) and were chemically stable and remained in solutionwithout precipitation (>48 hours).

The formulations of compound 12 from A (ii), A (iii) and A (iv) werediluted with D5W (10-fold) and remained in solution withoutprecipitation (>12 hours).

Example 53 Characteristics of Sodium Hydrochloride Citric Acid andTartaric Acid Salts or Complexes of Compound 12 Formulated in CAPTISOL

Sodium, hydrochloride, citrate and tartrate salts of a test compound ofFormula I were prepared in 30% CAPTISOL solutions and were studied forthe following:

Table G shows the physiochemical as well as pharmacokinetic (PK) andpharmacodynamic (PD) properties of sodium, hydrochloride, citric acidand tartaric acid salts of Compound 12.

TABLE G Sodium HCl Citric Acid Tartaric Acid Solubility 7.5 mg/ml 25mg/ml 25 mg/ml 60 mg/ml pH 10-11 2-3 4-5 3-4 IV Tissue High High LowHigh Dilution with D5W >10x >10x >10x >50x Chemical stability >12 h >12h >12 h >12 h in diluted solution 2-week chemical ND −20° C. ND −20° C.,stability in RT, 40° C. lyophilisate Deliverable highest 220-250 mg >750mg >750 mg >1800 mg daily dose in humans

Example 54 Comparison of Anti-Tumor Activity of Composition of Compound12 in 30% CAPTISOL and Erlotinib, a Prototype EGFRi in A549 NSCLCXenograft Model

Administration of compound 12 in 30% CAPTISOL attenuated tumor growth inthe NSCLC xenograft model. As shown in FIG. 11A, after 24 hours, animalstreated with compound 12 showed a 150% increase in tumor size whereasanimals treated with vehicle showed about a 240% change in tumor size.As shown in FIG. 11B, treatment of animals with Erolotinib did notsignificantly affect tumor size as compared to control.

Example 55 Effect of Composition of Compound 12 in 30% CAPTISOL in HPACPancreatic Cancer Cells

120 mg/kg of compound 12 in 30% CAPTISOL, 50 mg/kg erlotinib or vehiclewere administered to animals daily and change in tumor size over time(days) was measured. As shown in FIG. 12A, administration of 120 mg/kgcompound 12 in 30% CAPTISOL (iv/ip) resulted in greater attenuation oftumor growth than either erlotinib (po) or vehicle.

Pharmacokinetic Studies in Mice, Rats, and Dogs

The experimental methods used for Examples 77-83 are described below.

Animals: Mice (CD-1, male, 25-30 g), rats (Spraque Dawle, 260-300 g) anddogs (Beagles, male, 9-11 kg) were used for the PK studies. Animals wereprovided pelleted food and water ad libitum and kept in a roomconditioned at 23±1° C., humidity of 50-70%, and a 12-hour light/12-hourdark cycle.Drug Preparation and Administration. Compound 12 was dissolved in 30%CAPTISOL with equal molar concentration of tartaric acid or HCl orcitric acid, or NaOH. Compound was administered via an intravenous (iv)infusion. Conditions for iv infusion for each animal are shown below:

Mouse: 20 mg/kg and 60 mg/kg for 2 min i.v. infusioin

Rat: 20 mg/kg for 5 min i.v. infusion

Beagle: 25 mg/kg for 30 min i.v. infusion.

Blood and tissue Sample Collection. Blood was collected into tubescontaining sodium heparin anticoagulant at various time points. Theplasma was separated via centrifugation and stored in −40° C. beforeanalysis.Plasma Sample Extraction. Plasma samples were prepared by proteinprecipitation. An internal standard was added into plasma samples. A 50μl of plasma was combined with 150 μl of acetonitrile, vortexed, andcentrifuged for 10 min at 10000 rpm. The supernatant was then injectedonto LC/MS/MS.

Samples were compared to standards made in plasma. These standards wereprepared by serial dilution. An internal standard was added into theplasma with standard.

Tissue Sample Extraction. Lung and colon samples (20-200 mg) were usedfor extraction. Tissues were homogenized in 0.8 ml water. An internalstandard was added into the tissue homogenates. The homogenates wereextracted with 1-ml ethyl acetate for three times. After evaporation,the residual was reconstituted in 0.1 ml acetonitrile for LC/MS/MSassay.

LC/MS/MS Analytical Methods.

LC Conditions are Shown Below:

-   -   LC Instrument: Agilent HPLC 1100 Series    -   Autosampler: Agilent G1367A Autosampler    -   Analytical Column: YMC Pro C18 S3 (3μ, 2.0*50 mm, 120 Å)    -   Guard Column: YMC Pro C18 S3 Guard Column (3μ, 2.0*10 mm, 120 Å)    -   Column Temp: in ambient    -   Mobile Phase: A: acetonitrile:water:formic acid (5:95:0.1,        v/v/v)        -   B: acetonitrile:water:formic acid (95:5:0.1, v/v/v)    -   LC Gradient Program 0˜1 min: mobile phase A: 100%        -   1˜2.5 min: mobile phase A: 100% to 20%        -   2.5˜3 min: mobile phase A: 20%        -   3˜4 min: mobile phase A: 20% to 100%    -   Flow Rate: 200 μl/min    -   Autosampler Temp: in ambient    -   Injection Volume: 5 μl

Mass Spectrometer Conditions are Shown Below:

-   -   Instrument: PE Sciex API 3000    -   Interface: Turbo Ion Spray (TIS)    -   Polarity: Positive Ion    -   Scan: Multiple Reaction Monitoring (MRM)

Single or Multiple Dosing Toxicity Study in Mice and Rats

The experimental methods used for the toxicity study below are describedas follows:

Experiment Design:

1. Single dosing MTD in mice

-   -   a. CD-1 mice, male, 24-26 gram    -   b. Dosing at 0, 50, 100, 200, 400 mg/kg, iv infusion 2 min    -   c. 8 mice per group

2. Single dosing MTD in rats

-   -   a. Sprague Dawley, male and female, 240-260 gram    -   b. Dosing at 0, 25, 50, 100, 200 mg/kg, iv infusion 5 min    -   c. 6 rats per group (3 male and 3 female)

3. 7-day-multiple dosing MTD in mice

-   -   a. CD-1 mice, male, 24-26 gram    -   b. Dosing at 0, 50, 100, 200 mg/kg/d ip    -   c. 6 mice per group    -   d. Blood and organs will be collected 2 hr after last dosing on        Day 7 for hematology

4. 7-day multiple dosing MTD in rats

-   -   a. Sprague Dawley, male and female, 220-250 gram    -   b. Dose, 25, 50, 100, 200 mg/kg/d) iv infusion 5 min    -   c. 6 per group (3 male and 3 female)    -   d. Blood and organs will be collected 2 hr after last dosing on        Day 7 for hematology

Compound Preparation

The compound was dissolved in 30% Captisol with equal molarconcentration of tartaric acid. The stock solution: 25 mg/ml Tartaricform in 30% Captisol, 1 ml/vial store at −40° C. For example, 1000 mgcompound, 345 mg tartaric acid (0.345 mg tartaric acid per mg compound)and 40 ml 30% Captisol or 1000 mg compound, 2.3 ml 1N HCl (2.3 ul 1N HClper mg compound), 12 gram Captisol, Add water to 40 ml. Stock solutionis diluted with 30% CAPTISOL before use.

Example 56 Pharmacokinetics of Test Salts in Plasma, Lung and Colonafter Intravenous Administration

20 mg/kg of hydrochloride, citrate, sodium and tartrate salts ofcompound 12 in 30% CAPTISOL was administered intravenously to mice inorder to determine the concentration (ng/ml) over time (hours) ofcompound 12 after intravenous (iv) administration in plasma, lung andcolon. The results of these studies are shown in FIG. 13. As shownthere, the concentration of the tartrate salt of compound 12 in plasmadecreased from about 10,000 ng/ml in 2 hours. In lung, the concentrationof the tartrate salt of compound 12 decreased from about 10,000 ng/ml in6 hours. In colon, the concentration of the tartrate salt went decreasedfrom over 1000 ng/ml in 6 hours.

Example 57 Pharmacokinetic Study of Compound 12 Formulation in Mice

20 mg/kg and 60 mg/kg of a hydrochloride salt of compound 12 in 30%CAPTISOL was administered intravenously (iv) and intraperitoneally (ip)to mice and the half life (t½), maximal observed concentration (Cmax)and area under the curve (AUC) were determined. As shown in Table Hbelow, the concentration of compound 12 is dose proportional whenadministered intravenously but not intraperitoneally. The half-life ofcompound 12 in tissue is greater than that in plasma.

TABLE H Plasma Lung Colon 20 mgkg 60 mg/kg 20 mgkg 60 mg/kg 20 mgkg 60mg/kg IV Dose T ½ (hr) 0.2 0.3 3.9 1.9 1.7 2.2 Cmax (uM) 27.7 61.7 15.296.9 8.5 29.4 AUC (h*ng/ml) 715 3124 1571 8313 5529 13473 IP Dose T ½(hr) 0.26 0.51 2.2 3.5 NA NA Cmax (uM) 8.5 14.4 7.8 11.6 NA NA AUC(h*ng/ml) 3751 5721 4433 8309 NA NA

Example 58 Pharmacokinetic Study of Compound 12 Formulation in Rats

20 mg/kg and 60 mg/kg of a hydrochloride salt of compound 12 in 30%CAPTISOL was administered (iv) to rats and the concentration (ng/ml) ofthe compound was measured in plasma over thirty hours. As shown in FIG.14, the concentration of compound 12 in the plasma of the rat wasproportional to the dose of compound 12 administered.

Example 59 Single Dose IV Toxicity Study in Mice with the Compound 12Formulation

A single dose of compound 12 (25, 50, 100, 200 or 400 mg/kg) in 30%CAPTISOL was administered (iv) to mice and change in body weight wasmeasured over nine day to assess toxicity of the various doses ofcompound 12. As shown in FIG. 15, administration of up to 200 mg/kg ofcompound 12 did not result in a significant change in body weight.

Example 60 Seven Day Repeat IP Toxicity Study in Mice Using Compound 12Formulation

Repeated dosing of compound 12 over seven days (25, 50, 100, 200 or 400mg/kg) in 30% CAPTISOL was administered (ip) to mice and change in bodyweight was measured over seven days. As shown in FIG. 16, repeatedadministration of up to 100 mg/kg of compound 12 did not result in asignificant change in body weight.

Example 61 Single Dose IV Toxicity Study in Rats Using Compound 12Formulation

A single dose of compound 12 (25, 50, 100 or 200 mg/kg) in 30% CAPTISOLwas administered (iv) to rats and change in body weight was measuredover eight days to assess toxicity of the different doses of compound12. As shown in FIG. 17, administration of up to 200 mg/kg did notresult in a significant change in body weight.

The patent and scientific literature referred to herein establishes theknowledge that is available to those with skill in the art. All UnitedStates patents and published or unpublished United States patentapplications cited herein are incorporated by reference. All publishedforeign patents and patent applications cited herein are herebyincorporated by reference. All other published references, documents,manuscripts and scientific literature cited herein are herebyincorporated by reference.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A compound represented by formula I:

or a pharmaceutically acceptable salt, wherein X is O, S, CH₂, or —CONH—; B is a C₃ to C₉ alkylene; R₁ is selected from hydrogen, C₁ to C₄ alkoxy or substituted C₁ to C₄ alkoxy; R₂ is each independently selected from halogen, C₁ to C₄ alkyl, C₂ to C₄ alkenyl, and C₂ to C₄ alkynyl; and n is 1, 2 or
 3. 